Apparatus and methods relating to collecting and processing human biological material containing adipose

ABSTRACT

A portable apparatus for collection and processing of human biological material containing adipose, such as extracted during a lipoplasty procedure, is useful for multi-step processing to prepare a concentrated product (e.g., stromal vascular fraction) or a fat graft composition. The apparatus has a container with a containment volume with a tissue retention volume and a filtrate volume separated by a filter and with a tapered portion to a collection volume for collecting concentrate product. Inlet and suction ports provide access to the tissue retention volume and filtrate volume, respectively, and an extraction port provides versatile access for removal of target processed concentrate material or fat graft material, which access may be via a lumen through a rotatable mixer shaft. Access ports may be configured for access only from above the container. The apparatus is incorporatable into a variety of assemblies, systems, kits, methods and uses.

CROSS REFERENCE TO RELATED APPLICATIONS

This patent application claims priority to and thus the benefit of anearlier filing date from U.S. Provisional Patent Application No.61/363,150 (filed Jul. 9, 2010), the entire contents of which are herebyincorporated by reference.

FIELD OF THE INVENTION

The invention relates to apparatus, assemblies, systems, methods anduses related to collection and processing human biological materialcomprising adipose, including for preparation of fat grafts orconcentrate products such as stromal vascular fraction rich in stemcells.

BACKGROUND OF THE INVENTION

Adipose tissue is recognized as a promising source of stem cells with atleast multi-potent differentiation potential. Lipoasperate obtainedduring a lipoplasty procedure, such as lipo surgery, may be processed toprepare a so-called stromal vascular fraction (SVF) that is rich in stemcells. Processing to prepare SVF may include washing lipoasperate withsaline solution, followed by enzymatic digestion of washed tissue usingcollagenase, and centrifuging digested material to prepare SVF in theform of a centrifuged pellet. Such collection and processing of tissueinvolves several steps with transfer of contents between differentprocess containers for different tissue collection and processing steps,which is cumbersome and provides significant opportunities for error orcontamination.

Adipose is widely used in fat grafting, also referred to as fattransfer, procedures. There are a variety of different fat graftingapplications. One common fat graft application is breast augmentation orbreast reconstruction. Adipose is also a promising delivery vehicle, forexample to deliver stem cells or hormones with a fat graft. Currentpractices for collecting and processing adipose-containing tissue toprepare fat graft compositions do not adequately address needs acrossthe variety of applications in an efficient manner.

SUMMARY OF THE INVENTION

A first aspect of the invention is provided by an apparatus that isadapted to serve both for collection of human biological material (e.g.,tissue, biological fluids) comprising adipose removed from a patientduring a lipoplasty procedure and for convenient post-collectionprocessing of collected material. Such human biological material andseparated or processed portions thereof may be referred to herein simplyas “tissue”, for convenience and brevity. For example, the term tissuemay be used herein to refer to in-tact tissue, disrupted tissue, tissuefragments and biological fluids associated with or separate from tissue.The apparatus is orientable in a collection orientation for collectionof human biological material, or tissue, comprising adipose, such asduring a lipoplasty procedure. For convenience of description except asnoted, the apparatus is described as oriented in the collectionorientation. As such, relational references such as to top, bottom, up,down, above, below, elevations, vertical, horizontal and the like are inrelation to the apparatus as oriented in the collection orientation. Theapparatus may be configured such that the apparatus may be stablysupported in the collection orientation. For example, the apparatus mayhave a base configured for interfacing with a flat, substantiallyhorizontal surface (e.g., counter top or table top) to stably supportthe apparatus in the collection orientation, or may be held in amounting structure that maintains the apparatus in the collectionorientation. Although such an orientation is referred to as a“collection” orientation it should be appreciated that use of theapparatus is not limited to being oriented only in the collectionorientation or that only human biological material collection may beperformed while the apparatus is oriented in the collection orientation.The apparatus may be advantageously configured to permit performance ofmany different operations with the apparatus when the apparatus isoriented in the collection orientation.

The apparatus of the first aspect may be used in a variety of processingapplications involving adipose. The apparatus may, for example, be usedfor preparation of concentrated or separated portions of the collectedhuman biological material, for example to produce a stromal vascularfraction rich in stem cells derived from adipose tissue. As anotherexample, the apparatus may be used for preparation of a fat graftcomprising adipose. The apparatus has a design that accommodatesretention of a target material (e.g., stem cells or adipose) in a singlecontainer from collection through preparation of a concentrate or otherproduct containing the target material. By target material, it is meantsome component or components from or some portion or portions ofcollected human biological material of interest for recovery followingprocessing in the apparatus, such as recovery in a concentrated ormodified form relative to the collected human biological material (e.g.,stromal vascular fraction concentrate rich in stem cells, cleanedadipose-containing fraction for fat grafting applications)

The apparatus of the first aspect includes a container having aninternal containment volume, with the internal containment volumecomprising a tissue retention volume, a filtrate volume, a collectionvolume, and a tapered portion. The tissue retention volume and filtratevolume are separated by a filter with a separation size preferably in arange of from 70 microns to 400 microns. The collection volume is withinthe filtrate volume (i.e., is a part of the filtrate volume) and has abottom elevation corresponding to a bottom elevation of the filtratevolume and a top elevation that is lower than the bottom elevation ofthe tissue retention volume. The tapered portion tapers in a downwarddirection with at least a portion of the tapered portion being locatedabove the collection volume. The apparatus includes an inlet port influid communication with the tissue retention volume and configured forintroducing extracted biological material directly into the tissueretention volume, such as during a lipoplasty procedure. The apparatusincludes a suction port that is in fluid communication with the filtratevolume and provides access to the filtrate volume for suctioning fromthe filtrate volume material that may pass through the filter from thetissue retention volume to the filtrate volume, for example biologicalfluids that may separate from biological material introduced into thetissue retention volume.

As noted, the tissue retention volume and the filtrate volume of theapparatus of the first aspect are separated by the filter. By being“separated” by the filter it is meant that the filter forms at least aportion of the defining physical separation between the tissue retentionvolume and the filtrate volume. As will be appreciated, the filteraccommodates movement of fluid and undersize non-fluid material (e.g.,liberated undersize cells, tissue fragments, etc.) between the tissueretention volume and the filtrate volume, while retaining oversizematerial within the tissue retention volume. In some applications (e.g.,preparing a stromal vascular fraction concentrate), target material maycomprise biological material that during processing in the apparatuspasses through the filter and collects in the collection volume. Asdiscussed below, the apparatus may be configured to be received in acentrifuge for centrifuging the apparatus, and such centrifuging mayassist to concentrate such target material in the collection volume. Inother applications (e.g., preparing a fat graft), target material (e.g.,adipose) may be retained within the tissue retention volume followingprocessing.

The apparatus may be used during multiple processing steps to prepare,for example, a stromal vascular fraction from human biological materialcomprising adipose or a fat graft containing adipose, without the needto transfer a target material between different containers for differentprocessing steps. The apparatus may be used initially to collect thehuman biological material (e.g. tissue and fluids) during a lipoplastyprocedure. The apparatus may be portable and easily transportablebetween locations where collection or different processing operationsmay be conducted.

A number of feature refinements and additional features are applicableto the first aspect of the invention. These feature refinements andadditional features may be used individually or in any combination. Assuch, each of the following features may be, but are not required to be,used with any other feature or combination of the first aspect.

The apparatus may include an extraction port in fluid communication withthe internal containment volume and configured for removing processedbiological material from the internal containment volume. Any or all ofthe inlet port, the suction port and the extraction port may beconfigured for access therethrough from above the container into theinternal containment volume. The extraction port may be located above aportion of the filter, so that the advancing tip of a hypodermic needlepierces the filter when the tip of the hypodermic needle is advancedfrom the extraction port into the collection volume. The collectionvolume may include a nadir and the extraction port may be positionedabove the nadir so that the tip of a hypodermic needle inserted throughthe extraction port may be advanced vertically downward to the vicinityof the nadir of the collection volume.

The apparatus may include a mixing device disposed at least in partwithin the tissue retention volume for mixing contents within the tissueretention volume. The mixing device may comprise a rotatable shaftextending from outside of the internal containment volume to inside ofthe internal containment volume. The shaft may be made of a polymeric ormetallic material of construction. A preferred material of constructionfor the shaft is stainless steel, for example 303, 304 or 316 stainlesssteel. If a polymeric material of construction is used, it shouldpreferably be high-strength, for example an Ultem™ resin compositionavailable from Saudi Basic Industries Corp. (SABIC). The shaft maycomprise at the top a tapered receptacle adapted to mate with a taperedsyringe tip. The tapered receptacle may be fitted with an o-ring to sealagainst the tapered syringe tip when inserted into the taperedreceptacle, thereby permitting a good suction to be applied by thesyringe through the lumen to extract material from the tissue retentionvolume. The shaft may comprise a handle interface outside of theinternal containment volume. The apparatus may further include a handleinterfaced to the handle interface, wherein rotating the handle causesthe shaft to rotate, thereby operating the mixing device. The handle maybe removably interfaced to the handle interface, to permit the handle toconveniently be connected with the handle interface to operate themixing device when appropriate and to conveniently be removed from thehandle interface to permit access to a top of the apparatus withoutinterference from the handle. The apparatus may include a lumen thatextends through the shaft and has a proximal end located outside of theinternal containment volume and a distal end located within the internalcontainment volume. Such a lumen may provide access from outside of theinternal containment volume to inside of the internal containmentvolume. A removable plug for sealing the lumen may be disposed in theproximal end of the lumen. The shaft may be rotatable about an axis thatextends through the collection volume. The lumen may be aligned with theaxis. The lumen may provide access to the collection volume foraspiration of material therefrom and/or injection of material thereto.

In one variation on embodiments that include the lumen, the apparatusmay be configured for advancing a hypodermic needle through the lumenand out of the distal end of the lumen to access the collection volumewith an advancing tip of the hypodermic needle. The distal end of thelumen may be located in the tissue retention volume above a portion ofthe filter, so that the advancing tip of the hypodermic needle maypierce the filter when the tip of the hypodermic needle exits the distalend of the lumen and is advanced from the distal end of the lumen intothe collection volume. The collection volume may include a nadir, and anaxis of the lumen may be aligned so that the tip of a hypodermic needleexiting the distal end of the lumen may be advanced to the vicinity ofthe nadir of the collection volume. The hypodermic needle may thusaccess the collection volume to permit injection of material into and/oraspiration of material from the collection volume (e.g., aspiration ofstromal vascular fraction concentrate or other processed biologicalmaterial collecting in the collection volume during processing). Theapparatus may be designed for single-use, and piercing the filter with ahypodermic needle may beneficially provide a safety mechanism forpreventing reuse, and risks associated therewith, by damaging the filterin a way that renders the filter unsatisfactory for reuse.

In another variation on embodiments that include the lumen, a barriermember may be disposed between a distal end of the lumen and thecollection volume. The barrier member may thus prevent access throughthe lumen to the collection volume. In this way, the barrier member mayeffectively restrict use of the apparatus to applications in whichtarget material is retained within the tissue retention volume (e.g.,preparation of a fat graft), and may effectively prevent the apparatusfrom being used in applications in which target material may collect inthe collection volume. The barrier member improves the aspirationfunction through the lumen of the rotatable shaft by reducing oreliminating a potential pathway for air through the filter to the distaltip of the lumen from the filtrate volume to the distal tip of the lumenin the retention volume when air is present in the filtrate volume. Thedistal end of the lumen may be located in the tissue retention volumeand contents of the tissue retention volume may be removable from thetissue retention volume to outside of the internal containment volumethrough the lumen. Flow of the contents from the tissue retention volumeinto the lumen may be through a restricted space between the distal endof the lumen and the barrier member. In one preferred implementation,the distal end of the lumen and the barrier member may be separated by adistance of no more than 5 millimeters. The distal end of the lumen andthe barrier member may often, however, be separated by a distance of atleast 1 millimeter. When the lumen extends through a rotatable shaft,the barrier member may be connected with the shaft and may rotate whenthe shaft is rotated. The barrier member may be not connected to arotating shaft. For example, the barrier member may be part of orsupported by the filter or other structure disposed between the tissueretention volume and the filtrate volume.

The mixing device may comprise at one or more mixing members disposed inthe tissue retention volume and connected with the shaft, wherein themixing member moves through the tissue retention volume when the shaftis rotated. For example, a mixing member may be in the form of impellerblades, paddles or arms that agitate and mix components within thetissue retention volume when the shaft is rotated. At least a portion ofthe tissue retention volume may be within the tapered portion of theinternal containment volume, and at least a portion of one or more suchmixing member may be disposed within the tapered portion of the internalcontainment volume. The mixing device may include a filter contactmember that moves when the shaft is rotated and movably contacts thefilter. The filter contact member may be part of or separate from such amixing member. The filter contact member contacts the filter at leastperiodically, and may contact the filter continuously when the shaft isrotated. The filter contact member may advantageously deform the filterwhen it moveably contacts the filter, promoting dislodgment of materialfrom the filter to help prevent filter clogging.

As noted, the suction port is in fluid communication with the filtratevolume. By the suction port being in fluid communication with thefiltrate volume, it is meant that the suction port is fluidly connecteddirectly to the filtrate volume, and not indirectly through the tissueretention volume and the filter. The fluid communication may be providedby a dedicated conduit extending from the suction port to a desiredlocation within the filtrate volume where it is desired to apply suctiondirectly to the filtrate volume. The suction port may be in fluidcommunication with the tapered portion of the internal containmentvolume through a conduit providing fluid communication from the suctionport to a location within the filtrate volume that is also within thetapered portion of the internal containment volume. The conduit mayextend through the filtrate volume from adjacent the suction port tosuch a location within the filtrate volume. The suction port may belocated above the tapered portion of the internal containment volume.The suction port may be configured for access through the suction portfrom above the container. The suction port may be configured forconnection to a vacuum system to suction material from the filtratevolume, such as material that passes through the filter from the tissueretention volume to the filtrate volume.

The apparatus may include multiple suction ports. For example, theapparatus may include a first suction port as described in the precedingparagraph that is in fluid communication with a first location in thefiltrate volume within the tapered portion of the internal containmentvolume through a first conduit, and the apparatus may include a secondsuction port through which components passing through the filter fromthe tissue retention volume to the filtrate volume may be suctioned fromthe filtrate volume through a second conduit extending from the secondsuction port to a second location within the filtrate volume. The secondconduit may be configured to permit adjustment of the elevation of thesecond location within the filtrate volume. The second conduit may betranslatable through the second suction port to adjust the elevation ofthe second location within the filtrate volume. The second conduit maybe configured so that at any extent of such adjustment of the secondlocation, the second location will always be at a higher elevationwithin the filtrate volume than the first location, which may be fixed.The second conduit may be configured to permit adjustment of theposition of the second location within the filtrate volume at differentelevations above the tapered portion of the filtrate volume. The secondconduit may be configured to permit positioning the second location atan elevation corresponding with an interface between the tapered portionof the filtrate volume and a portion of the filtrate volume locatedabove the tapered portion, which may be the lowest elevation to whichadjustment is permitted. The second suction port may be configured foraccess through the second suction from above the container.

Any one or more of the inlet port, the suction port of other portsproviding access to the internal containment volume may be configuredfor access through the port from above. In this way, access through eachsuch port may be conveniently from above the apparatus, providing asignificant advantage to a user of the apparatus in that such a user mayfocus all access manipulations from above the apparatus while theapparatus is in a normal position in the collective orientation, forexample with the apparatus freestanding on a flat work surface such as atable or counter. Complexities associated with access from the side orfrom below may be avoided, including the complexity of sealing andproviding access into side or bottom access ports against a positivefluid head that may be present in the container and the complexity ofawkward side of bottom interactions by users. Although such access fromabove the container may be at some angle relative to vertical, in apreferred implementation the access through such port is in a verticaldirection from above the container. In one preferred implementation, allaccess to the internal containment volume may be through access portswherein each such access port (e.g., inlet port, suction port,extraction port, other ports) is configured for access through theaccess port only from above the container. In another preferredimplementation, all access ports may be configured for access througheach such access port in a vertical direction from above the container.

The tapered portion may have a cross-sectional area that tapers, orreduces in size, in a direction toward the bottom of the collectionvolume. The tapered portion of the containment volume helps to directand concentrate target dense material (e.g., dense cells, stromalvascular fraction) toward and into the collection volume. The taperedportion of the containment volume may have a conical shape or any othershape with a cross-sectional area that tapers to reduce in size in adirection toward the bottom of the collection volume.

The container may have a self-supporting structure, with the containerhaving rigid walls or rigid structural supports to maintain thecontainer in a particular configuration. The apparatus may have acontainer that is not self-supporting. The container may have anon-self-supporting wall structure. For example, the container may havea wall structure formed by a flexible vinyl or other plastic bagmaterial. Such a non-self supporting wall structure (e.g., plastic fluidcontainment bag) may be sealed to a rigid top of the container, whichtop may comprise one or more of the inlet port, the suction port and/orother ports.

The container may be made of any suitable material or materials ofconstruction. The container may be made of one or more plasticcomposition. The container may have transparent walls. A preferredmaterial of construction for the container is a clear polymericmaterial, such as for example a clarified polypropylene composition.Clarified polypropylene compositions provide low cellular adhesion andreasonable clarity. The container may be comprised of multiple pieces,which may all be made out of the same material of construction or one ormore of such pieces may be made of a different material of construction.

The container may comprise a fluid containment shell with an internalcavity portion forming at least a part of the internal containmentvolume. The internal cavity portion may be open to above. The containermay include a lid attached to the shell and disposed to cover from abovethe internal cavity portion. One or more than one of the suction portthe inlet port or other access port may pass through the lid. In onepreferred implementation, all access into the internal containmentvolume may be only through one or more openings, or ports, passingthrough the lid. The filter may be suspended from the lid. The mixingdevice may be supported by the lid and extend vertically downward fromthe lid into the tissue retention volume. The apparatus may include aflow barrier skirt extending between 5 mm and 50 mm downward from thelid into the internal containment volume. The shell may comprise wallsaround the internal cavity portion except where the cavity portion isopen to above, and the apparatus may be configured with no access intothe internal containment volume through the walls of the shell. Theshell may comprise an upper portion having a first wall surface portiondefining a corresponding upper portion of the internal containmentvolume. Substantially all of the first wall surface portion may have asteep incline relative to horizontal, for example an incline of at least65°, preferably an incline of at least 75° and more preferably anincline of about 90° (vertical wall surface). The shell may include alower portion located below the upper portion and having a second wallsurface portion defining a corresponding lower portion of the internalcontainment volume, and the second wall surface portion may include atapered wall surface portion defining the tapered portion of theinternal containment volume. The tapered wall surface portion may have aless steep incline relative to horizontal than the first wall portion ofthe upper portion. The incline relative to horizontal of the taperedwall portion, or of the entire second wall portion when comprisedentirely of the tapered wall portion, may be in a range having an upperlimit of 70°, 65°, 60° or 65° and a lower limit of 20°, 25°, 30° or 35°,with one preferred range being form 30° to 60°. The tapered portion ofthe internal containment volume may occupy substantially the entirelower portion of the internal containment volume, and the second surfacemay be made up entirely or substantially entirely by the tapered wallsurface. At least a first portion of the filter may be disposed in theupper portion of the internal containment volume and a second portion ofthe filter may be disposed in the lower portion of the internalcontainment volume. The incline of each of the first wall surfaceportion, the second wall surface portion and the tapered wall surfaceportion need not be uniform, however all portions of the first wallsurface portion may preferably be at a steeper incline than the inclineof any portion of the tapered wall surface portion.

There are a number of advantages that may be available withconfigurations of the preceding paragraph including an upper portionhaving a steeper wall surface incline and a lower portion having a lesssteep wall surface incline. Such a structure advantageously accommodatesa larger proportion of the internal containment volume being allocatedto the tissue retention volume, with a larger portion of the tissueretention volume in the upper portion of the shell and a smaller portionof the tissue retention volume in a lower portion of the shell. Thetapered wall surface portion of the lower portion of the shell helps todirect fluid and other material in the filtrate volume toward the bottomof the filtrate volume for efficient collection and removal. Inapplications where target material is to be collected in the collectionvolume, the tapered wall surface also directs material toward thecollection volume, which may generally be located in a bottom portion ofthe filtrate volume. The apparatus may also be configured to becentrifugable, and centrifuging will tend to accelerate concentration ofa most dense fraction (e.g., stromal vascular fraction) in thecollection volume.

More generally, the tapered portion of the internal containment volumemay have a tapered portion nadir corresponding with a bottom elevationof the internal containment volume. The bottom elevation of thecollection volume may correspond with the bottom elevation of theinternal containment volume. Wall surfaces of the container defining thetapered portion of the internal containment volume may coverage at apoint at the tapered portion nadir. This is a particularly beneficialconfiguration, especially for applications when target material is to becollected in and removed from the collection volume in the vicinity ofthe tapered portion nadir.

The apparatus may be configured with a very convenient size from anumber of perspectives, and with efficient use of the internalcontainment volume to facilitate efficient collection of biologicalmaterial and versatility in post-collection processing. The apparatusmay be sized for convenient hand transportation, such as between alocation where human biological material may be collected to other,different locations, where various processing of collected material maybe carried out. The apparatus may also be sized for convenientmanipulation by a person.

For many applications, the apparatus may be sized and configured suchthat the internal containment volume has a volume in a range with alower limit of 100 cubic centimeters, 300 cubic centimeters, 500 cubiccentimeters, 600 cubic centimeters or 700 cubic centimeters and an upperlimit of 1500 cubic centimeters, 1300 cubic centimeters, 1100 cubiccentimeters, 1000 cubic centimeters, 900 cubic centimeters or 800 cubiccentimeters. One preferred range for many applications is for theinternal containment volume to be in a range of 700 cubic centimeters to1000 cubic centimeters. By internal containment volume, it meant thetotal internal volume contained within the walls defining the container,including volume that is occupied by internal hardware, such as forexample may be occupied by a mixing device, barrier member, suctionconduits, barrier skirt, etc. As will be appreciated, less than all ofthe internal containment volume will be available for processing withinthe internal containment volume.

The terms “available processing volume” or “useful volume” are usedinterchangeably herein to refer to the portion of the internalcontainment volume that is effectively available to receive and processhuman biological material and additives (e.g. wash other additives)during use of the apparatus for collection of biological material or forpost-collection processing. This available processing volume is equal tothe internal containment volume less portions of the internalcontainment volume occupied by hardware (e.g., mixing device, filter,skirt, suction tubes, barrier member, etc) and less unoccupied portionsof the internal containment volume not effectively accessible foroccupation by biological material during collection operations or bybiological material or additives during post-collection processing. Forexample, the available processing volume may exclude a small volume atthe top of the container that is above a bottom extension of the inletport into the internal containment volume. This small void space may bebeneficial to permit space for fluid to slosh within the container whencontents of the container may be internally mixed or externally agitated(e.g., by a shaker table). For many applications, the availableprocessing volume may be in a range having a lower limit of 75 cubiccentimeters, 200 cubic centimeters, 400 cubic centimeters, 500 cubiccentimeters, 600 cubic centimeters, 650 cubic centimeters or 700 cubiccentimeters and an upper limit of 1300 cubic centimeters, 1100 cubiccentimeters, 1000 cubic centimeters, 900 cubic centimeters, 850 cubiccentimeters, 800 cubic centimeters or 750 cubic centimeters. In onepreferred implementation for many applications, the available processingvolume may be in a range of from 700 cubic centimeters to 850 cubiccentimeters.

Advantageously, the apparatus may be configured so that a large portionof the available processing volume is within the tissue retentionvolume, while still permitting a high level of performance for variousprocessing operations. The tissue retention volume may comprise at least60 percent, at least 65 percent or at least 70 percent of the availableprocessing volume with the container. Often, the tissue retention volumewill comprise not more than 95 percent, not more than 90 percent or notmore than 85 percent of the available processing volume. For manypreferred implementations, the tissue retention volume may comprise aportion of the available processing volume that is at least 400 cubiccentimeters, at least 500 cubic centimeters, at least 600 centimeters orat least 650 cubic centimeters. The apparatus may advantageously beconfigured with only a small portion of the available processing volumeoccupied by the collection volume, located below the filter. Forexample, the collection volume may comprise no more than 10 percent, nomore than 7 percent or no more than 5 percent of the availableprocessing volume.

For many preferred implementations the collection volume may be nolarger than 75 cubic centimeters, no larger than 50 cubic centimeters,no larger than 30 cubic centimeters or no larger than 20 cubiccentimeters. In one preferred implementation, the collection volume maybe in a range of from 10 cubic centimeters to 30 cubic centimeters.Typically, the entire collection volume will make up part of theavailable processing volume.

The apparatus may be sized and configured to be containable within arelatively small envelope volume which may be particularly advantageousgiven the relatively large internal containment volume, availableprocessing volume and tissue retention volume that may be provided inthe container. For some preferred implementations, the apparatus may besized and configured to be containable within a first envelope volumedefined by a rectangular cuboid having a length dimension of no morethan 16 centimeters, a depth dimension of no more than 15 centimetersand a height dimension of no more than 18 centimeters. However, theapparatus may be sized and configured to have some minimum size, forexample as a function of a desired size of internal containment volume,available processing volume or tissue retention volume.

For some preferred implementations, the apparatus may have a size andconfiguration such that the apparatus may not be containable within asecond envelope volume defined by a rectangular cuboid having any one ormore of a length dimension, depth dimension or height dimension that issmaller than 10 centimeters, i.e., the apparatus would not fit withinany rectangular cuboid smaller than 10×10×10 centimeters.

The filter may be of any appropriate filter media design. The filter maybe any porous structure with openings sized to make a desiredseparation. The filter may be in the form of a mesh filter bag disposedwithin the internal containment volume, and that separates the internalcontainment volume between the tissue retention volume and the filtratevolume located on opposite sides of the filter bag. The filter may be arigid mesh screen. In one implementation, the filter may have aseparation size in a range having a lower limit of 70 microns andpreferably 80 microns and an upper limit of 125 microns, preferably 110microns and more preferably 100 microns. By separation size, it is meantthe size at which the filter effects separation between particlespassing through and particles rejected by the filter during normaloperation. The separation size may be determined by the size of openingsprovided in a surface filter, such as the mesh size of a mesh bag filteror of a rigid mesh screen filter.

In one preferred implementation, the filter may be a mesh filter. With amesh filter, the separation size will be the size of the mesh openings.In one preferred implementation, whether or not the filter is a meshfilter, the separation size for the filter, and the size of meshopenings when a mesh filter is used, may be in a range having a lowerlimit of 70 microns, 80 microns, 90 microns, 100 microns, 125 microns or150 microns and having an upper limit of 400 microns, 350 microns, 300microns or 250 microns. A particularly preferred range for manyapplications is from 150 microns to 250 microns, including forpreparation of a fat graft or preparation of a stromal vascular fractionconcentrate. The mesh filter may be of a flexible or a rigid meshmaterial. In a preferred implementation, the filter may be made of meshmaterial, more preferably a nylon mesh material. The filter need not becontinuous, and may be comprised of discrete filter areas disposed atdifferent locations between the tissue retention volume and the filtratevolume. Alternatively, the filter may be, comprised of a singlecontinuous filter area. The filter defines at least part of the physicalseparation between the tissue retention volume and the filtrate volume;it need not define all of the physical separation between the issueretention volume and the filtrate volume. For example, there may beinternal walls defining at least a part of the physical separationbetween the tissue retention volume and the filtrate volume, with anexample being a skirt barrier that may be disposed at the top of theinternal containment volume and that may define a separation between thetissue retention volume and the filtrate volume in an upper portion ofthe internal containment volume. Another example may be a barrier memberthat blocks access from the tissue retention volume into the collectionvolume portion of the filtrate volume. As another example, the filtermay include filter areas supported by a frame, with structural membersof the frame defining apart of the physical separation between thetissue retention volume and the filtrate volume. In a preferredimplementation, the portion of the physical separation between thetissue retention volume and the filtrate volume that is provided thefilter should generally be large to provide as much filter surface areaas reasonably possible.

The apparatus may be configured to be received by a centrifuge forcentrifuging. For example, the apparatus may be conveniently sized andconfigured to be received within a centrifuge bucket, and preferably ofa commercially available centrifuge. For example, the apparatus mayadvantageously be sized and configured to fit within a bucket of abucket assembly for a Sorvall ST-40 centrifuge. Two or more of theapparatus may be centrifuged simultaneously in a centrifuge. In onepreferred implementation, the apparatus may be sized and configured sothat two of the apparatus may be simultaneously centrifuged together ina dual bucket assembly, wherein each apparatus counterbalances the otherapparatus during centrifuging, for efficient processing. Alternativelyone apparatus could be processed within a blank weight as acounterbalance. The apparatus may be received in a centrifuge bucketwith the bottom of the apparatus adjacent to and facing the bottom ofthe bucket.

Configuring the apparatus for centrifuging may be particularlyadvantageous for applications when target material is to be collected inthe collection volume, such as for collecting a stromal vascularfraction concentrate in the collection volume. The collection volume isadvantageously positioned in the bottom portion of the internalcontainment volume where the most dense materials will collect duringcentrifuging.

The apparatus may include an extraction port in direct fluidcommunication with the filtrate volume and through which material isremovable from the filtrate volume separate from the suction port. In apreferred implementation, access through the extraction port is invertical direction from above the container, for example with theextraction port passing through the top of the container. Although not apreferred implementation, the apparatus may include an extraction portthat is adjacent a bottom elevation of the filtrate volume andconfigured for access to the collection volume. Such an extraction portmay comprise a sealing mechanism that is normally sealed and selectivelymanipulable to extract material from the collection volume. The sealingmechanism may comprise a septum sealably penetrable by an extractionneedle. The collection volume may comprise a collection chamber withinthe filtrate volume adjacent to the extraction port, which chamber isconfigured so that the apparatus may be centrifuged to collect withinthe collection chamber at least the most dense components present withinthe filtrate volume during a centrifuging operation. The apparatus maycomprise a stromal vascular fraction from adipose tissue, which may bedisposed within the collection chamber. The tapered portion of theinternal containment volume may be adjacent to the collection chamberand opposite the extraction port. The tapered portion of the containmentvolume may help to direct and concentrate target dense cells toward andinto the collection chamber. The collection chamber may be cylindricallyshaped, preferably with sufficient volume within the cylinder toaccumulate all target concentrate product (e.g., all of a stromalvascular fraction). The extraction port may be manipulable toselectively extract material that has collected in the collectionchamber. The extraction port may include a sealing mechanism that isnormally sealed to normally retain material in the collection chamberand that is manipulable to extract material from the collection chamberas desired. Such a sealing mechanism may include, for example, a septumor diaphragm sealably penetrable by an extraction needle (e.g., ahypodermic needle) or a valve. The extraction port may comprise aneedleless port for engaging the tip of a syringe. Preferably, theextraction port is located immediately adjacent the bottom of thecollection chamber. The container may have a top side and a bottom side,with the extraction port adjacent to the bottom side and with the inletport adjacent to the top side. The suction port may be adjacent to thetop side of the container and in fluid communication with a conduitextending from adjacent the top side to adjacent the collection chamberto permit, for example, suction of components from within the filtratevolume in the vicinity of the collection chamber.

The apparatus may be configured for the addition of additives to theinternal containment volume, and in particular directly into the tissueretention volume. As used herein, such an additive is any material addedto the internal containment volume other than the human biologicaltissue comprising adipose, such as from a lipoplasty procedure, to beprocessed in the apparatus. Such an additive may be added for example toaid processing within the apparatus or to become part of a compositionincluding target material to be recovered from the apparatus forfollowing processing. Example of some additives to aid processing mayinclude wash liquid, enzymes or surfactants. Examples of some additivesthat may become part of a fat graft composition include hormones (e.g.,human growth hormone, insulin), buffers (e.g., sodium bicarbonate) andcells (e.g., bone marrow-sourced stem cells, cultured adipose-sourcedstem cells, stromal vascular cells from adipose tissue). Such additivesmay be added to the tissue retention volume through the inlet port.

The apparatus may further include a second inlet port in fluidcommunication with the tissue retention volume for introducing anadditive directly into the tissue retention volume. The second inletport may be smaller than the inlet port through which the humanbiological material to be processed (e.g., from a lipoplasty procedure)is introduced into the tissue retention volume. The second inlet port,which may be referred to as an additive inlet port, may be convenientlysized and configured for insertion therethrough of a hypodermic needlefrom which an additive material may be ejected from the needle into thetissue retention volume.

The apparatus may include human biological material, which may includetarget material from human biological material originally collected inthe apparatus disposed within the internal containment volume (e.g., theapparatus during some stage of use). The apparatus may includeadipose-containing material in the tissue retention volume. Theapparatus may include in the tissue retention volume anadipose-containing fat graft composition, including any desiredadditives, ready to be withdrawn from the tissue retention volume andused in a fat graft procedure. The apparatus may include a stromalvascular fraction concentrate disposed in the collection volume.

The apparatus may be packaged within a hermetic enclosure, for exampleas packaged for transportation and storage prior to use. The apparatusmay be sterilized prior to packaging and maintained in a sterileenvironment within the hermetic enclosure at least until the apparatusis removed from the hermetic enclosure for use. The apparatus may bedesigned for a single use following removal from the hermetic enclosure.After such single use, the apparatus may be disposed of in anappropriate manner.

The apparatus may be fluidly connected through the inlet port to apre-filter. The fluid connection may be through a conduit that fluidlyconnects the inlet port to a downstream side of a pre-filter unitcomprising the pre-filter. The pre-filter unit may comprise a housingwith the pre-filter disposed within the housing, and with an inlet on anupstream side of the pre-filter and an outlet on downstream side of thepre-filter. The upstream side of the pre-filter may be in fluidcommunication with a lipoplasty cannula to receive extracted biologicalmaterial during a lipoplasty procedure and to pre-filter the biologicalmaterial prior to delivery to the inlet port of the apparatus, forexample to remove some collagen from the biological material upstream ofthe apparatus. The pre-filter may be provided with my convenientseparation size, for example in a range having a lower limit of 0.5millimeter or 1 millimeter to an upper limit of 5 millimeters, 3millimeters or 2 millimeters. The pre-filter may comprise a mesh screenwith mesh openings sized, for example, in such a range.

A second aspect of the invention is provided by an assembly comprising acombination of an apparatus according to the first aspect and at leastone other device. A number of feature refinements and additionalfeatures are applicable to the second aspect of the invention. Thesefeature refinements and additional features may be used individually orin any combination. As such, each of the following features may be butare not required to be, used with any other feature or combination ofthe second aspect or the first aspect.

The assembly may comprise a rigid mounting structure, such as a rigidshell, to support the apparatus, or the container of the apparatus, forexample during use of the apparatus for collecting human biologicalmaterial or for post-collection processing. The rigid mounting structuremay be made, for example, of metal or plastic material of construction.Combination of the apparatus with a rigid mounting structure isparticularly useful when the container comprises a non-self-supportingwall structure, wherein the container is supported by the mountingstructure in a configuration suitable for a human biological materialcollection or processing operation, for example with the non-selfsupporting wall structure maintained in a fully extended and operationalconfiguration. In one implementation of the assembly, the container maycomprise a top from which the non-self supporting wall structure issuspended, and which top is supported by the rigid mounting structure.The non-self-supporting wall structure may be sealed to the top. Such anon-self-supporting wall structure may be provided in the form of a bag,such as made from a vinyl or other plastic material. The centrifuge maybe a centrifuge as described above in relation to the first aspect.

In one implementation, the assembly may comprise the apparatus and acentrifuge. The apparatus may be received in the centrifuge forCentrifugation of the apparatus, and also any contents in the apparatus.In one variation, the assembly comprises at least two of the apparatusreceived in the centrifuge for simultaneous centrifugation, and in onevariation the assembly comprises precisely two of the apparatus receivedin the centrifuge. In one preferred variation, each such apparatus isreceived in the centrifuge in a manner so that the collection chamber ofthe internal containment volume will collect at least the most densematerial within the filtrate volume during centrifuging of theapparatus, and will also collect the desired concentrate product,whether or not that is comprised of the densest material.

In one implementation, the assembly may comprise the apparatus and anagitation device. The apparatus may be mounted on the agitation devicefor agitation of the apparatus, and also any contents of the apparatus.The agitation device may be, for example, a shaker (e.g., shaker table).In one variation, the agitation device may be a shaker with temperaturecontrol capability for controlling the temperature of the apparatus andcontents, and preferably for warming and maintaining the apparatuscontents at a constant temperature. In one implementation, each suchapparatus, as mounted on the agitation device, is at a controlledtemperature of from 35-39° C., and preferably at about 37° C. Theagitation device may include a warmer-shaker. The apparatus mounted onthe agitation device may include human biological material comprisingadipose or a component derived from human biological material comprisingdisposed within the internal containment volume.

A third aspect of the invention is provided by a tissue collection andprocessing system comprising the apparatus fluidly connected to receiveor discharge fluid or other components to or from the internalcontainment volume of the container. A number of feature refinements andadditional features are applicable to the third aspect of the invention.These feature refinements and additional features may be usedindividually or in any combination. As such, each of the followingfeatures may be, but are not required to be, used with any other featureor combination of the first aspect or the third aspect.

The tissue collection and processing system may comprise the apparatusfluidly connected through the inlet port to a tissue conduit forconducting human biological material comprising adipose to the inletport for introduction into the tissue retention volume of the container.This might be the case, for example, during a lipoplasty procedure withthe tissue conduit connected to a liposuction cannula through whichhuman biological material comprising adipose is removed from a patientduring the lipoplasty procedure. The tissue collection and processingsystem may comprise the apparatus fluidly connected through the suctionport to a vacuum system for applying suction to the filtrate volume.This may be the case, for example, during a lipoplasty procedure whenthe apparatus is receiving human biological material comprising adiposeinto the tissue retention volume, and material passing through thefilter during the lipoplasty procedure being removed from the filtratevolume during the lipoplasty procedure by the suction applied throughthe suction port. In one implementation, the suction port may be fluidlyconnected to the vacuum system through a collection canister forcollecting fluid suctioned from the filtrate volume, which fluid maycomprise components of the human biological material that pass throughthe filter from the tissue retention volume to the filtrate volume, suchas may occur during a lipoplasty procedure. Such fluid suctioned fromthe filtrate volume may, for example, comprise red blood cellsseparating from the human biological material and pulled through thefilter and into the filtrate volume by suction applied through thesuction port. The collection canister may, for example, be a wastecanister to collect waste products for disposal.

The tissue collection and processing system may comprise the apparatus,a vacuum system, a tissue feed conduit, and a suction conduit. Thevacuum system may be in fluid communication with the suction port of theapparatus and may be capable of applying suction to the filtrate volumeof the apparatus. The tissue feed conduit may be in fluid communicationwith the inlet port of the apparatus to conduct human biologicalmaterial comprising adipose into the tissue retention volume through theinlet port. The suction conduit may be in fluid communication with thesuction port and the vacuum system to suction fluid from the filtratevolume of the apparatus. The suction port may be in fluid communicationwith the vacuum system through a collection canister to collect fluidsuctioned from the filtrate volume through the suction port. The tissuefeed conduit may be in fluid communication with a lipoplasty cannula.The tissue feed conduit may be in fluid communication with thelipoplasty cannula through a pre-filter to pre-filter human biologicalmaterial prior to introduction into the tissue retention volume of theapparatus. The pre-filter may be as described with respect to the firstaspect, and may be in a pre-filter unit that is or includes features asdescribed in relation to the first aspect. The tissue collection andprocessing system may include human biological material extracted duringa lipoplasty procedure moving through the tissue feed conduit forintroduction of human biological material into the tissue retentionvolume, and a vacuum applied to the filtrate volume by the vacuum systemthrough the suction conduit and drawing into the suction conduit fluidcomprising components passing through the filter from the tissueretention volume.

A fourth aspect of the invention is provided by a method for processingtissue from a lipoplasty procedure within a containment volume of aportable tissue collection and processing apparatus to prepare withinthe apparatus a concentrate product comprising at least one targetcomponent of the tissue. The apparatus comprises a filter and acontainer having an internal containment volume, wherein the internalcontainment volume comprises a tissue retention volume and a filtratevolume separated by the filter. The method comprises washing the tissuewith a wash liquid, with the washing comprising adding the wash liquidto the internal containment volume to contact tissue within the tissueretention volume and passing through the filter and removing from thefiltrate volume at least a portion of the wash liquid along with one ormore component, typically a non-target component, washed from the tissuewhile retaining washed tissue within the tissue retention volume. Afterthe washing, the method comprises digesting tissue within thecontainment volume. The digesting comprises adding enzyme to thecontainment volume to contact at least a portion of washed tissue withinthe tissue retention volume. The enzyme is of a type capable of breakingdown a portion of the washed tissue to release the target component in aform capable of passing through the filter, and which may then berecovered through the filtrate volume. The digesting comprises, afteradding the enzyme, agitating the contents of the containment volume fora time and at a temperature sufficient to release the target componentin such a form capable of passing through the filter. After thedigesting, the method comprises centrifuging the apparatus to prepare inthe filtrate volume a concentrate product comprising the targetcomponent.

A number of feature refinements and additional features are applicableto the fourth aspect of the invention. These feature refinements andadditional features may be used individually or in any combination. Assuch, each of the following features may be, but are not required to be,used with any other feature or combination of the first aspect, thesecond aspect or the third aspect.

The portable tissue collection and processing apparatus may be anapparatus according to the first aspect of the invention. The method mayinclude one or more steps in addition to the washing, digesting andcentrifuging steps. Any such additional step may be performed eitherprior to the washing, between the washing and digesting, between thedigesting and centrifuging, or after the centrifuging. The agitatingperformed during the digesting may be performed, for example, using anassembly of an apparatus and agitation device according to the secondaspect of the invention. The agitating may be performed by or mayinvolve use of a mixing device disposed within the internal containmentvolume. The centrifuging may be performed, for example, using anassembly of an apparatus received in a centrifuge according to thesecond aspect of the invention.

In one implementation, the washing may comprise a multi-stage operationwith multiple occurrences of adding wash liquid followed by removingused wash liquid from the filtrate volume. The washing may comprise,after adding the wash liquid to the containment volume, centrifuging theapparatus prior to the digesting to help separate wash liquid fromtissue that is retained in the tissue retention volume. Following thecentrifuging, wash liquid collecting in the filtrate volume may beremoved from the filtrate volume prior to the digesting, such as forexample through the suction port and/or through the extraction port.When the washing comprises multiple washing stages, each such stage maycomprise adding wash liquid to the containment volume followed bycentrifuging the apparatus. The washing may be in the absence of suchcentrifuging. The washing may comprise, after adding the wash liquid,mixing contents (including the wash liquid) in the tissue retentionvolume (e.g., by operating a mixer in the tissue retention volume),discontinuing the mixing after some period of time and suctioning thefluids from the filtrate volume. The washing may comprise multiplestages with multiple occurrences of adding wash liquid, mixing,discontinuing mixing and suctioning. Following discontinuing the mixing,some quiescent time may be allowed for decantation of liquid prior tocommencing the suctioning. The wash liquid may comprise any suitableliquid for washing the tissue within the tissue retention volume. Thewash liquid may be or comprise a saline solution, for example aphosphate buffer solution. The saline may be warmed, for example toapproximately normal human body temperature, prior to introduction intothe tissue retention volume.

The enzyme added to the containment volume during the digestingpreferably includes collagenase, which may be accompanied by a neutralprotease. The agitating may be accomplished by shaking the apparatusmounted on a shaker, preferably with control of temperature, such as ona warming shaker. The agitating may comprise mixing contents within allor part of the internal containment volume through operation of a mixingdevice disposed in the internal containment volume, for example arotating mixing member (e.g., blade, impeller), preferably with controlof temperature. During the agitating, the apparatus may be maintained ata relatively constant temperature, for example, between about 35° C. and39° C., and preferably around 37° C.

The method may comprise prior to the washing, collecting the tissue inthe tissue retention volume of the container. Collecting the tissue maybe accomplished during a lipoplasty procedure (e.g., lipo surgery) on apatient, such as by conducting adipose tissue that is being removed fromthe patient into the tissue retention volume through a conduit fluidlyconnected with the apparatus. The tissue collection may be accomplished,for example, with a tissue collection and processing system according tothe third aspect of the invention. The collecting may comprise, duringsuch a lipoplasty procedure, suctioning at least a portion of fluidcollecting in the filtrate volume to remove such fluid from, thefiltrate volume and from the apparatus. Such fluid may comprise liquidand/or small particles that pass through the filter from the tissueretention volume into the filtrate volume during the lipoplastyprocedure. Such fluid may be removed from the filtrate volume by suctionthrough the suction port. Applying suction to the internal containmentvolume has the benefit of pulling filterable components through thefilter into the filtrate volume for removal. Removing such fluid fromthe filtrate volume permits the filtrate volume to be kept small becauseit is not necessary to contain all fluid that may separate from theadipose tissue and pass through the filter during a lipoplastyprocedure. This provides the advantage of allowing the tissue retentionvolume to be made proportionally larger to collect a larger quantity ofdesired tissue during a tissue collection operation. Applying suction tothe filtrate volume aids separation of fluid components from the adiposetissue, further providing an advantage of reducing the portion of tissueretained and allowing a larger quantity of desired tissue to becollected during the collection operation.

The method may comprise after the collecting and prior to the washing,disconnecting the apparatus from the conduit through which it receivedadipose tissue, sealing the containment volume and transporting theapparatus from a first location where the collecting is performed to asecond, different location where the washing is performed. The methodmay comprise after the centrifuging, selectively removing theconcentrate product from the filtrate volume. For example, when using anapparatus of the first aspect of the invention, the concentrate productmay collect in a collection volume, which may be in a collection chamberat the bottom of the collection volume. The concentrate product may beand may be selectively removed from the collection volume through anextraction port. By “selectively removing”, it is meant that theconcentrate product is removed separate from one or more other materialspresent in the filtrate volume following the centrifuging. For example,the filtrate volume may contain multiple different material phases ofdifferent densities within the filtrate volume, whether or not multiplephases are present in the collection volume. The targeted concentrateproduct may be one or more of these phases. During the selectivelyremoving, the desired phase or phases representing the concentrateproduct are removed separate from some or all of the other phases thatmay be present in the filtrate volume. In one preferred implementationof the method, a target component comprises stem cells from adiposetissue. The concentrate product may comprise a stromal vascularfraction, which may be in a cell fraction collecting at the bottom ofthe collection volume. In the case in particular of a stromal vascularfraction in the concentrate product, there may be a phase rich in redblood cells that collect in the collection volume below, and possiblygrades into, the bottom of the stromal vascular fraction. There may alsobe one or more less dense fractions above the stromal vascular fractionwithin the filtrate volume, some of which may also occupy a portion ofthe collection volume above the stromal vascular fraction. In oneimplementation, such a red blood cell phase may be removed from thefiltrate volume, for example through the extraction port, prior toremoval of the stromal vascular fraction, when the extraction port isadjacent the bottom of a collection chamber. The red blood cell phasemay be removed from the filtrate volume together with the stromalvascular fraction in a combined cell fraction. Through thorough washingduring the washing operation, the red blood content in such aconcentrated cell fraction may be kept relatively small, such that thestromal vascular fraction may be utilized directly in the recoveredconcentrated cell fraction containing the relatively small red bloodcell content. If desired, after removal from the apparatus such a redblood cell phase may be separated from the stromal vascular fraction byknown techniques. In one implementation, the concentrate product, suchas containing a stromal vascular fraction, is in the form of a pellet,and to facilitate removal of the concentrate product, the concentrateproduct is diluted in the filtrate volume with a diluent fluid and adiluted concentrate product is then removed from the filtrate volume.The diluent fluid may comprise or be a saline solution, such as forexample a phosphate buffer solution. Prior to removing the stromalvascular fraction, one or more less dense layers may first be removedfrom above the stromal vascular fraction, such as via a suction port.

A fifth aspect of the invention is provided by a method for collectinghuman biological material comprising adipose extracted from a humanpatient during a lipoplasty procedure. The method includes introducing afeed comprising human biological material with adipose into a tissueretention volume within an apparatus, retaining a retained portion ofthe feed in the tissue retention volume, and passing a separated portionof the feed through a filter from the tissue retention volume to afiltrate volume within the apparatus. The introducing the feed includespassing the feed through an inlet port into the tissue retention volume.

A number of feature refinements and additional features are applicableto the fifth aspect of the invention. These feature refinements andadditional features may be used individually or in any combination. Assuch, each of the following features may be, but are not required to be,used with any other feature or combination of the any of the firstthrough fifth aspects.

The apparatus may an apparatus according to the first aspect of theinvention.

The method may include, during at least a portion of the introducing,removing from the filtrate volume through the suction port at least aportion of the separated portion of the feed. The method may comprisediscontinuing the introducing and performing the removing for someperiod of time after the discontinuing the introducing. During theremoving, substantially all of the separated portion of the feed may beremoved from the filtrate volume through the suction port.

In an embodiment, the volume of the separated portion of the feed may belarger than the available processing volume within the filtrate volume.In some implementations, the total volume of feed introduced into thetissue retention volume during the introducing may be greater than theavailable processing volume within the tissue retention volume. Forexample, the total volume of feed introduced into the tissue retentionvolume during the introducing may be at least 125 percent or at least150 percent of available processing volume within the tissue retentionvolume. The retained portion of the feed may comprise from 25 to 75volume percent of the feed introduced into the tissue retention volumeduring the introducing. The retained portion of the feed may occupy avolume of 50 cubic centimeters, at least 200 cubic centimeters, at least400 cubic centimeters or at least 500 cubic centimeters within thetissue retention volume. The feed may comprise human biological materialpassing through a pre-filter located upstream of the inlet port to thetissue retention volume.

A sixth aspect of the invention is provided by a method for processinghuman biological material comprising adipose within an internalcontainment volume of an apparatus. The method includes washing thehuman biological material with a wash liquid. The washing includesadding the wash liquid to the internal containment volume to contact thehuman biological material within a tissue retention volume within theinternal containment volume. Within the internal containment volume, afilter separates the tissue retention volume form a filtrate volume. Thewashing further includes passing through the filter and removing fromthe filtrate volume at least a portion of the wash liquid along with oneor more components washed from the human biological material whileretaining washed human biological material in the tissue retentionvolume.

A number of feature refinements and additional features are applicableto the sixth aspect of the invention. These feature refinements andadditional features may be used individually or in any combination. Assuch, each of the following features may be, but are not required to be,used with any other feature or combination of any of the first throughsixth aspects.

The apparatus may be an apparatus according to the first aspect of theinvention.

The washing may be according to the washing described with respect tothe fourth aspect. For example, the washing may or may not involvecentrifuging after adding wash liquid, and may involve a single washingstage or may involve a multi-stage washing operation.

The method may further include adding an additive to the washed humanbiological material in the tissue retention volume. The method mayfurther include mixing the washed human biological material to dispersethe additive in the washed human biological material. The mixing mayinclude moving a member within the tissue retention volume. The additivemay comprise an ingredient for delivery to a patient in a fat graft. Theadditive may, for example, be any of the additives listed for possibleinclusion in a fat graft composition as discussed in relation to thefirst aspect.

The method may include removing from the tissue retention volume atleast a portion of the washed human biological material for use in a fatgraft.

The method may include digesting material within the containment volume.The digesting material may include adding enzyme to the containmentvolume to contact at least a portion of the washed human biologicalmaterial within the tissue retention volume. Such an enzyme may be of atype capable of breaking down a portion of the washed human biologicalmaterial to release a target component in a form capable of passingthrough the filter. Such an enzyme may comprise collagenase, orcollagenase with neutral protease. The digesting may include, after theadding enzyme, agitating contents of the containment volume for a timeand at a temperature sufficient to release the target component in theform capable of passing through the filter. The method may include,after the digesting, centrifuging the apparatus to prepare in thecollection volume a concentrate product comprising the target component.The target component may include stem cells from adipose tissue. Theconcentrate may comprise a stromal vascular fraction. The method maycomprise, after the centrifuging, selectively removing the concentrateproduct from the filtrate volume. After the centrifuging and prior tothe selectively removing, the concentrate product may be contained in abottom separated layer or layers disposed within the collection volume,and the selectively removing may include removing from the filtratevolume other separated layers within the filtrate volume disposed abovethe bottom separated layer or layers containing the concentrate product.When the apparatus comprises a second suction port, the removing fromthe filtrate volume such other separated layers within the filtratevolume may include removing material of the other separated layersthrough the second suction port.

The removing from the filtrate volume other separated layers within thefiltrate volume disposed above the bottom separated layer or layerswithin the concentrate product may include tilting the container andsuctioning material of the other, higher layers from a laterally distantlocation within the filtrate volume that is laterally distant from anouter edge of the concentrate product. During such suctioning theconcentrate product may remain in a pellet located adjacent the bottomof the collection volume. The laterally distant location may be adjacenta top edge of the tapered portion of the internal containment volume.The internal containment volume may include a corner at the top edge ofthe tapered portion, and the laterally distant location may be adjacentthe corner.

After removing from the filtrate volume separated layers within thefiltrate volume disposed above the bottom separated layer or layers withthe concentrate product, the method may include diluting such bottomlayer or layers with a diluent fluid to prepare a diluted concentrateproduct and removing the diluted concentrate product from the filtratevolume. The other separated layers removed from above the concentrateproduct may comprise an aqueous layer, a disaggregated adipose layer(containing remnants of disaggregated adipose tissue) and an oil layer.The concentrate product may comprise a concentrated cell fractioncontaining stromal vascular cells (stromal vascular fraction) and a redblood cell fraction.

The digesting may include shaking the apparatus mounted on a shakerwhile maintaining the apparatus at a controlled elevated temperature.The digesting may be or include in features as described for digestingin relation to the fourth aspect.

The method may comprise, prior to the washing, collecting the humanbiological material. The collecting the human biological material mayinclude, during a lipoplasty procedure on a patient, conducting a humanbiological material feed comprising at least a portion of extractedhuman biological material removed from the patient into the tissueretention volume. The collecting may include, during the lipoplastyprocedure, passing fluid separating from the human biological materialfeed through the filter into the filtrate volume and suctioning at leasta portion of the fluid from the filtrate volume and out of theapparatus. After the collecting and prior to the washing, the method mayinclude sealing the containment volume and transporting the apparatusfrom a first location where the collecting is performed to a second,different location where the washing is performed.

A seventh aspect of the invention is provided by a kit that includes anapparatus according to the first aspect of the invention and apre-filter unit comprising a pre-filter with openings sized for sizeseparation in a range of from 0.5 mm to 5 mm.

A number of feature refinements and additional features are applicableto the seventh aspect of the invention. These feature refinements andadditional features may be used individually or in any combination. Assuch, each of the following features may be, but are not required to be,used with any other feature or combination of the first, second, thirdor seventh aspects.

The kit may further include a pre-filter conduit having a first endconfigured to be connected to an outlet port of the pre-filter and asecond end configured to be connected to the inlet port of theapparatus. The pre-filter may comprise a mesh screen with mesh openingsin a range having a lower limit of 0.5 millimeter or millimeter and anupper limit of 5 millimeters, 3 millimeters or 2 to 2 millimeters. Theapparatus and the pre-filter unit may be contained within a commonpackaging container. The pre-filter conduit may also be contained withthe common packaging container. The common packaging container maycomprise a box. In an arrangement, each of the apparatus and thepre-filter unit may be sterilized and disposed within a hermeticenclosure. The pre-filter unit may be or have features as described fora pre-filter unit in relation to the first aspect of the invention.

An eighth aspect of the invention is provided by a uses, or methods ofusing, the apparatus of the first aspect of the invention.

A use, or method of using, may be for processing human biologicalmaterial comprising adipose. A use, or method of using, may be forpreparation of a fat graft. A use, or method of using, may be forpreparation of a stromal vascular fraction concentrate.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows in perspective a tissue collection and processingapparatus.

FIG. 2 shows the same tissue collection and processing apparatus as FIG.1 with some component parts shown in exploded view.

FIG. 3 shows in perspective a tissue collection and processing apparatusmounted on a rigid mounting structure.

FIG. 4 shows in perspective the same tissue collection and processingapparatus as shown in FIG. 3, removed from the rigid mounting structure.

FIG. 5 shows the same tissue collection and processing apparatus asshown in FIGS. 3 and 4, with some component parts shown in explodedview.

FIG. 6 shows in perspective a tissue collection and processing system.

FIG. 7 shows side and top view of the same tissue collection andprocessing system as shown in FIG. 6.

FIG. 8 shows side and end sectional views of a tissue collection andprocessing apparatus having collected tissue disposed therein.

FIG. 9 shows side and end sectional views of a different tissuecollection and processing apparatus, supported by a rigid mountingstructure and having disposed therein collected tissue.

FIG. 10 shows in perspective a tissue collection and processingapparatus having collected tissue disposed therein, and with inlet andsuction ports capped.

FIG. 11 shows two tissue collection and processing apparatus beingloaded into a centrifuge for centrifuge processing.

FIG. 12 shows two tissue collection and processing apparatus mounted inrigid mounting structures being loaded into a centrifuge for centrifugeprocessing.

FIG. 13 shows a tissue collection and processing apparatus being loadedinto a centrifuge for centrifuge processing.

FIG. 14 shows two tissue collection and processing apparatus loaded in aspinning centrifuge.

FIG. 15 shows a tissue collection and processing apparatus suspended bya handle and from which material is being removed from the filtratevolume through an extraction port.

FIG. 16 shows the same tissue collection and processing apparatus asshown in FIG. 15 during removal of material from the filtrate volumethrough an extraction port.

FIG. 17 shows a tissue collection and processing apparatus suspended bya handle and from which material is being removed from the filtratevolume through a suction tube inserted into a suction port.

FIG. 18 shows a tissue collection and processing apparatus being mountedon a warmer-shaker.

FIG. 19 shows a tissue collection and processing apparatus being mountedon a warmer-shaker.

FIG. 20 shows a centrifuge with two tissue collection and processingapparatus received therein for centrifuge processing.

FIG. 21 shows a tissue collection and processing apparatus suspended bya handle and from which material is being removed from a collectionchamber through an extraction port.

FIG. 22 shows top, perspective, side and end views of another embodimentof a tissue collection and processing apparatus.

FIG. 23 shows another perspective view of the same tissue collection andprocessing apparatus as FIG. 22.

FIG. 24 shows the same tissue collection and processing apparatus asFIG. 23 with a shell removed.

FIG. 25 shows the same tissue collection and processing apparatus asFIG. 24 with a filter removed.

FIG. 26 illustrates various regions within the tissue collection andprocessing apparatus of FIG. 23.

FIG. 27 shows a configuration of the tissue collection and processingapparatus of FIG. 23.

FIG. 28 shows another configuration of the tissue collection andprocessing apparatus of FIG. 23.

FIG. 29 illustrates a needle inserted into a tissue collection andprocessing apparatus.

FIGS. 30A and 30B illustrate a translatable conduit in a tissuecollection and processing apparatus.

FIG. 31 illustrates a pre-filter and a tissue collection and processingapparatus.

FIGS. 32 through 35 illustrate steps in a method of processing tissuewithin a tissue collection and processing apparatus.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS

FIG. 1 shows a tissue collection and processing apparatus 100 of adesign designated for convenience as “Design A”. FIG. 2 shows the sameapparatus 100 as in FIG. 1, but illustrated in an exploded view of someof the components. As shown in FIGS. 1 and 2, the apparatus 100 has amesh bag filter 102 disposed within an internal containment volume of acontainer. The container is comprised of a rigid shell 104 and a rigidtop 106. The top 106 is sealed to the top of the shell 104. The shell104 may be made, for example, from a transparent plastic composition.The lid 106 may be made, for example, from a plastic composition.Disposed at the bottom of the shell 104 is a collection chamber 108 andan extraction port 110. The extraction port 110 comprises a sealingmechanism that is manipulable to extract material from inside thecollection chamber 108. The sealing mechanism may, for example, comprisea silicon rubber septum or diaphragm that may be penetrable by ahypodermic needle to extract material, or may comprise a valve.

The mesh filter bag 102 divides and separates the internal containmentvolume of the container into a tissue retention volume 112 disposedinside the mesh filter bag 102, and a filtrate volume 113 disposedwithin the shell 104 on the outside of the mesh filter bag 102. Thefiltrate volume 113 is that portion of the internal containment volumeinto which filtrate enters after passing through the mesh filter bag 102from the tissue retention volume 112.

An inlet port 114 in fluid communication with the tissue retentionvolume 112 through the top 106 is configured for introducing adiposetissue directly into the tissue retention volume during a lipoplastyprocedure, such as for example through a tissue transport conduit thatmay be attached to the inlet port 114 to conduit tissue from a patientduring such a procedure. An additional access port 116 in fluidcommunication through the top 106 with the tissue retention volume 112provides an additional route for introducing material into or removingmaterial from the tissue retention volume 112. A suction port 118 is influid communication through the top 106 with the filtrate volume 113 viaa conduit 120 extending from the suction port 118 to the vicinity of thetop of the collection chamber 108. The suction port 118 is configuredfor connection to a vacuum system, for example through connection of asuction conduit through which suction may be applied by a vacuum systemto suction from the filtrate volume material passing through the meshfilter bag 102 from the tissue retention volume 112 into the filtratevolume 113. The shell 104 has a tapered wall portion 122 that defines atapered portion of the internal containment volume, such that thecross-sectional area of the tapered portion of the internal containmentvolume tapers with a reducing cross-sectional area in a direction towardthe collection chamber 108. By tapering, it means that thecross-sectional area in a horizontal plane (assuming the apparatus 100is in an upright position as shown in FIG. 1) becomes smaller in acontinuous manner in the direction of the taper (e.g., a directionorthogonal to the horizontal plane). The collection chamber 108comprises a cylindrical volume located immediately below the taperedportion of the internal containment volume. The cross-sectional area ofthe internal containment volume at the bottom of the tapered portion 122approaches that of the cross-sectional area of the collection chamber108. “Collection chamber 108” is used to refer both to the downwardlyextending cylindrically-walled portion of the shell body 104 and thecylindrical portion of the filtrate volume disposed therein.

The shell 104 includes a base portion 123 configured to support theapparatus 100 in the upright position as shown in FIG. 1, for examplewhen the apparatus is resting on top of a hard flat surface, such as atable or shelf. For convenience, the side of the container adjacent thetop 106 is referred to as the top side of the container and the side ofthe container adjacent the extraction port 110 is referred to as thebottom side of the container.

The apparatus 100 comprises mixers 124 that have agitator arms 126 thatare rotatable to help mix contents within the internal containmentvolume, and in particular within the tissue retention volume 112. Themixers 124 may be driven by electrical power to rotate the agitator arms126, which power may be supplied, for example, by an external electricalpower source or by batteries disposed within the body of the mixers 124or elsewhere in the apparatus 100.

The apparatus 100 includes a retractable handle 128 to facilitatesuspending the apparatus 100 or for grasping and holding the apparatus100 by hand. As shown in FIG. 1, the handle 128 is in an extendedposition for use to grasp or suspend the apparatus 100. FIG. 2 shows thehandle 128 in a retracted position that is conveniently out of the wayso that the handle 128 does not interfere with access to the inlet port114, the access port 116 or the suction port 118 during use of theapparatus 100.

The apparatus 100 is designed to be portable, and is preferably portableby someone grasping the handle 128 and picking up the apparatus 100 bythe handle 128 by hand, preferably by using a single hand, to facilitateready transport of the apparatus 100, either while the internalcontainment volume is empty or with human tissue or components thereofdisposed within the internal containment volume.

FIGS. 3-5 illustrate a tissue collection and processing apparatus 140 ofa design designated for convenience as “Design B”. The apparatus 140 ofFIGS. 3-5 has a container with a non-self-supporting wall structure, asdescribed further below, and is shown in FIGS. 3-5 in combination with arigid mounting structure 142 on which the apparatus 140 may be mountedto accommodate the non-self-supporting wall structure, for example, whenthe apparatus 140 is being used for collection of tissue during alipoplasty procedure or at other times when it is desired to providesupport to the apparatus 140. The features of the apparatus 140 are bestseen in FIGS. 4 and 5, where features of the apparatus 140 are shownseparated from the rigid mounting structure 142.

With continued reference to FIGS. 3-5, the apparatus 140 has a containerformed by a rigid top 144 and a flexible containment bag 146 that issealed to and suspended from the top 144. Inside the container is aninternal containment volume that is separated into a tissue retentionvolume 148 and a filtrate volume 150 by a mesh filter bag 152. Disposedat the bottom of the filtrate volume is a collection chamber 154. Accessto the collection chamber 154 is provided through an extraction port 156having a sealing mechanism that is manipulable to extract material fromthe collection chamber 154. The apparatus 140 has an inlet port 158 influid communication with the tissue retention volume 148, an access port160 also in communication with the tissue retention volume 148, and asuction port 162 in fluid communication with the filtrate volume 150 viaa conduit 164 extending to the vicinity of the top of the collectionchamber 154. The containment bag 146, in a fully extended andoperational configuration as shown in FIGS. 3-5 has a tapered wallportion 166 corresponding with a tapered portion of the internalcontainment volume, wherein the cross-sectional area of the internalcontainment volume is tapered to reduce in size toward the top of thecollection chamber 154. A retractable handle 168 facilitates suspensionand hand grasping of the apparatus 140.

The top 144, mesh filter bag 152, collection chamber 154, extractionport 156, inlet port 158, access port 160, suction port 162, conduit164, and handle 168 as shown for the apparatus 140 of FIGS. 3-5 may, forexample, be of the same or similar design as the top 106, mesh bagfilter 102, collection chamber 108, extraction port 110, inlet port 114,access port 116, suction port 118, conduit 120 and handle 128 of theapparatus 100 shown in FIGS. 1 and 2 and described above.

A difference between the Design A of the apparatus 100 shown in FIGS. 1and 2 and the Design B of the apparatus 140 shown in FIGS. 3-5 is thatthe container of the apparatus 140 shown in FIGS. 3-5 is defined belowthe top 144 by the containment bag 146, whereas the apparatus 100 shownin FIGS. 1 and 2 has a containment volume defined below the top 106 bythe walls of the rigid shell 104. The containment bag 146 of theapparatus 140 shown in FIGS. 2-5 provides a less expensive alternativefor providing the desired containment volume, which may be importantespecially when the device is designed for a single use application.However, because the walls of the containment bag 146 are notself-supporting, and because the containment bag 146 would be morefragile than the rigid shell 104, the rigid mounting structure 142 maybe provided both to support the apparatus 140 and to provide protectionto the walls of the containment bag 146 during at least some uses of theapparatus 140. The design for the rigid mounting structure 142 shown inFIGS. 3-5 has a shape and design similar to the rigid shell 104 shown inFIGS. 1 and 2, but the rigid mounting structure 142 is not a part of theapparatus 140, but rather is an auxiliary device that may be used incombination with the apparatus 140.

The containment bag 146 may be made, for example, from a vinyl or otherflexible plastic material. The shell 142 may be made, for example, froma rigid plastic composition such as may be used for the rigid shell 104of FIGS. 1 and 2.

The apparatus 140 is shown in FIGS. 3-5 is not fitted with internalmixers corresponding with the mixers 124 of the apparatus 100 as shownin FIGS. 1 and 2. However, if desired, the same or other mixer devicescould be incorporated into the design of the apparatus 140 of FIGS. 3-5.

Reference is now made to FIGS. 6 and 7 showing a tissue collection andprocessing system 200 including the tissue collection and processingapparatus 100 of Design A with an inlet port fluidly connected to atissue conduit 204. A suction port of the apparatus 100 is fluidlyconnected with a canister 206 via a suction conduit 208. The canister206 is fluidly connected with a vacuum system (not shown) through aconduit 210. For illustration purposes, the apparatus 100 is of a typeof Design A. However the apparatus of Design B or of any other designcould be used in such a tissue collection and processing system. Duringoperation of the tissue collection and processing system 200, the tissueconduit 204 is conducting adipose tissue to the inlet port of theapparatus 100 for introduction of the adipose tissue into the tissueretention volume of the apparatus 100. Suction is applied to thefiltrate volume within the apparatus 100 by the vacuum system throughthe conduit 210, the canister 206 and the suction conduit 208 to removeby suction from the filtrate volume material separating from the adiposetissue in the tissue retention volume of the apparatus 100 and passingthrough the filter and into the filtrate volume of the apparatus 100.Such material suctioned from the filtrate volume through the suctionconduit 208 may then be collected in the canister 206. The canister 206may be a waste canister and the collected material may be waste forappropriate disposal. For example, red blood cells may separate fromadipose tissue during collection of the adipose tissue in the apparatus100 and such red blood cells passing through the filter will be removedfrom the filtrate volume of the apparatus 100 by suction via the suctionconduit 208.

In a method for processing tissue from a lipoplasty procedure, thetissue may be processed within a containment volume of a portable tissuecollection and processing apparatus to prepare within the apparatus aconcentrated product comprising at least one target component, or atleast one target material, from the tissue. The apparatus has a filterand a container having an internal containment volume, wherein theinternal containment volume comprises a tissue retention volume and afiltrate volume separated by the filter. The apparatus may, for example,be a tissue collection and processing apparatus of the Design A, DesignB or some other appropriate design. The method may comprise: washingtissue in the containment volume with a wash liquid; after the washing,digesting tissue within the containment volume; and after the digestion,centrifuging the apparatus to prepare in the filtrate volume aconcentrate product comprising at least one target component. Forexample the concentrate product may comprise, or may consist essentiallyof, stromal vascular fraction from adipose tissue, and a targetcomponent may be or comprise stem cells from adipose tissue. The methodmay also comprise one or more steps in addition to the washing,digesting and centrifuging. For example such an additional step mayoccur prior to the washing, between the washing and digesting, betweenthe digesting and centrifuging or after the centrifuging.

During the washing, the wash liquid may be added to the containmentvolume to contact tissue within the tissue retention volume and with atleast a portion, preferably a majority, and more preferably most, of thewash liquid passing through the filter into the filtrate volume. Thewash liquid may wash one or more component from the tissue whileretaining washed tissue in the tissue retention volume. The washedtissue may be retained in the tissue retention volume by the filter.Wash liquid passing into the filtrate volume may be removed from thefiltrate volume, along with any component or components washed from thetissue. After adding the wash liquid, an optional step of centrifugingthe apparatus may be performed. Such centrifuging may facilitate a highdegree of separation of the wash liquid from the tissue retained in thetissue retention volume. Next, the wash liquid may be removed from thefiltrate volume, for example by being suctioned through a suction portof the apparatus or by removal through an extraction port of theapparatus. The wash liquid may be an aqueous liquid, and may be orcomprise a saline solution, for example a phosphate buffer solution. Toensure thorough washing of the tissue, the washing may include multiplewash stages, with each stage comprising adding wash liquid to thecontainment volume to contact tissue within the tissue retention volumeand removing wash liquid from the filtrate volume.

During the digestion, an enzyme, such as for example collagenase, isadded to the containment volume to contact at least a portion,preferably a majority of, and more preferably all or substantially allof the washed tissue, within the tissue retention volume. The enzymeshould be of a type capable of breaking down at least a portion of thewashed tissue to an extent to release a target component, or material,in a form capable of passing through the filter and into the filtratevolume. After adding the enzyme, the digesting may comprise agitatingcontents of the containment volume of the apparatus for a time and at atemperature sufficient for the digestion to proceed to an extent tosignificantly release the target component, or material, in the desiredform capable of passing through the filter. The agitating may involveany method to agitate contents of the containment volume, including forexample one or both of: (a) shaking the apparatus to agitate thecontents within the apparatus and (b) mixing the contents within theapparatus, such as with one or more mixing device disposed within thecontainment volume and preferably disposed within the tissue retentionvolume. Shaking the apparatus may be accomplished by mounting theapparatus on a shaker, and preferably a warmer-shaker with a temperaturecontrol feature so that the apparatus and its contents may be maintainedat a controlled temperature, such as at or approximately at human bodytemperature.

Post-digestion centrifuging promotes separation of the target componentfrom the digested tissue and passage of the target component through thefilter for collection in the filtrate volume, such as for example tocollect within a collection chamber at the bottom of the apparatus, forexample a collection chamber such as shown in FIG. 1 or FIG. 2. Thecentrifuging causes a concentrate product to collect in the filtratevolume, and preferably in such a collection chamber. Multiple materialphases may collect within the filtrate volume, one or more of which orone or more portions of which, may comprise the desired concentrateproduct containing a target component.

Before the washing, the method may comprise collecting the tissue in theinternal containment volume of the apparatus. The collecting maycomprise conducting adipose tissue removed from a patient during alipoplasty procedure into the tissue retention volume through a tissueconduit fluidly connected with the apparatus during the lipoplastyprocedure. Such collection may be performed, for example, using a tissuecollection and processing system such as shown in FIGS. 6 and 7. Duringthe collecting, fluid separating from the adipose tissue and passingthrough the filter into the filtrate volume may be immediately removedfrom the filtrate volume by suctioning the fluid from the filtratevolume and out of the apparatus, for example to a collection canistersuch as that shown in FIGS. 6 and 7.

The method may comprise, after the centrifuging, removing theconcentrate product from the filtrate volume of the apparatus. Theconcentrate product may, for example, be removed from an apparatus suchas those illustrated in FIGS. 1 and 2 from the collection chamber at thebottom of the filtrate volume through the extraction port withmanipulation of the sealing mechanism. During the removing, theconcentrate product may be removed to and collected in the barrel of asyringe, or in the barrels of multiple syringes. The removing mayinclude selectively removing the concentrate product from the filtratevolume, to isolate the concentrate product from other material that mayhave collected in the filtrate volume during the centrifuging. To assistremoving the concentrate product, the concentrate product may be dilutedwith a dilution liquid to put the concentrated product in a dilute formthat is easier to remove from the filtrate volume. The dilution liquidmay be an aqueous liquid. The dilution liquid may be a saline solution,for example a phosphate buffer solution. Such dilution of theconcentrate product may be particularly useful in the situation wherethe concentrate product collects in the form of a relatively hard pelletin the filtrate volume, which may be the case for collection of stromalvascular fraction from adipose tissue. Selective removal of theconcentrate product may include separating a material phase comprisingthe concentrate product from one or more other material phase thatcollects in the filtrate volume. For example, a stromal vascularfraction may be a middle density phase, with a more dense phase and aless dense phase disposed on either side of the stromal vascularfraction. The more dense phase, which may collect at the bottom of acollection chamber in the apparatus, may be rich in red blood cells.Selective removal of the stromal vascular fraction may include firstremoving this red blood cell phase from the collection chamber (e.g.,into a syringe) and then removing the stromal vascular fraction from thecollection chamber (e.g., into a different syringe).

The apparatus used with the method for processing tissue may be portableand versatile, such as for example is the case with the apparatus ofDesign A or Design B. The method may include transporting the apparatus,and tissue contained therein, between locations where differentprocedures are performed. For example, the apparatus may be located atone location where collecting tissue is performed, while the washingand/or digesting may be performed at a different location. In oneimplementation, the containment volume comprising tissue may be sealedfollowing one procedure and the apparatus with the sealed containmentvolume may be transported to a different location for performance of asubsequent procedure. With the apparatus of Design A or Design B asdescribed previously, the apparatus may be sealed by capping the inletport, access port and suction port with sealing caps.

To further illustrate various features of the method for processingtissue, and apparatus, assemblies and systems that may be used duringthe method, reference is now made to FIGS. 8-21.

FIGS. 8 and 9 show the apparatus 100 of Design A (FIG. 8) and theapparatus 140 of Design B (FIG. 9) during a tissue collecting operation.During tissue collection, adipose tissue from a lipoplasty procedure,which may be referred to as lipoasperate, is received in the tissueretention volume 112 or 148. FIGS. 8 and 9 show material 224 or 226 thatseparate from lipoasperate, pass through the filter 102 or 152 andcollect in the respective filtrate volume 113 or 150, while tissue 220or 222 is retained in the respective tissue retention volume 112 or 148.The material 224 or 226 collecting in the respective filtrate volume 113or 150 may be continuously or intermittently removed from the filtratevolume 113 or 150 by suction through the respective suction port 118 or162 and the respective conduit 120 or 164.

FIG. 10 shows an example of the apparatus 140 of Design B followingtissue collection with the inlet port 158 and the suction port 162sealed by sealing caps 230. The access port 160 is left open for accessinto the tissue retention volume to remove materials from or to addmaterials to the tissue retention volume. For example, a light oil phasethat may develop on top of the collected tissue may be removed throughthe access port 160. As another example, wash liquid may be addedthrough access port 160 for washing collected tissue.

FIGS. 11, 12 and 13 each shows two of the apparatus 100 of Design A(FIG. 11) or two of the apparatus 140 of Design B (FIG. 12 and FIG. 13)being loaded into centrifuge buckets 240 of a two-bucket centrifugerotor and bucket assembly 242 of a centrifuge 244. As shown in FIG. 12,the apparatus 140 of Design B is being loaded into the centrifuge whileit remains mounted in the rigid mounting structure 142, while in FIG. 13the apparatus 140 of Design B is shown being loaded into the centrifugebuckets 240 in the absence of such a rigid mounting structure. Suchcentrifuging may be preformed for example as an option as part of awashing operation to help separate wash liquid from tissue or during thecentrifuging step after digestion for collection of the concentrateproduct in the filtrate volume.

FIG. 14 shows two of the apparatus 100 of Design A received in thecentrifuge buckets 240 of the two-bucket rotor and bucket assembly 242of centrifuge 244 while the centrifuge 240 is operating to spin theassembly 242 to centrifuge the apparatus 100 during a centrifugingoperation.

FIGS. 15 and 16 show the apparatus 100 of Design A suspended by thehandle 128 following optional centrifuging of a washing operation andprior to digesting with a hypodermic needle 250 inserted through asealing mechanism 252 in the extraction port 110 for removal ofinfranatant (e.g., dirty wash liquid) from the filtrate volume 113 andinto the barrel of a syringe 254.

FIG. 17 shows the apparatus 100 of Design A following optionalcentrifuging of a washing operation and prior to digestion, in whichinfranatant (e.g., dirty wash liquid) is being removed from the filtratevolume 113 via a suction tube 260 inserted through the suction port 118and the conduit 120 into the filtrate volume 113, with the materialremoved from the filtrate volume 113 being collected within the barrelof a syringe 262.

FIGS. 18 and 19 show the apparatus 100 of Design A (FIG. 18) and theapparatus 140 of Design B (FIG. 19) being mounted on a warmer-shaker 270for controlled temperature agitation of the contents of the respectiveapparatus 100 or 140, such as may be performed during a digestionoperation after adding an enzyme to the internal containment volume. Inthe case of the apparatus 100 of Design A, the mixers within theinternal containment volume may be operated during the shaking operationto assist agitation in addition to the shaking provided by thewarmer-shaker 270. With the apparatus 140 of Design B, the apparatus 140may be agitated in the absence of a rigid mounting structure 142. Toassist agitation that may be provided by the shaking motion of thewarmer-shaker, the warmer-shaker may also include paddles that operateagainst flexible walls of the containment bag 146.

FIG. 20 shows two of the apparatus 100 of Design A received in thecentrifuged 244 for centrifuging, as may be optionally performed duringa washing operation or as may be performed during the centrifugingfollowing digestion.

FIG. 21 shows the apparatus 140 of Design B suspended from the handle168 and with a concentrate product being selectively removed from thecollection chamber 154 through a sealing mechanism in the extractionport 156 through a hypodermic needle 288 into the barrel of a syringe290. For example, the concentrate product being selectively removed asshown in FIG. 21 may be a stromal vascular fraction from adipose tissue.

FIG. 22 shows an apparatus 300 for collection of tissue comprisingadipose removed from a patient during a lipoplasty procedure and forpost-collection processing of collected tissue. The apparatus 300 isillustrated in a collection orientation. The collection orientation isthe orientation in which the apparatus 300 may be placed during thecollection of adipose removed from a patient during a lipoplastyprocedure. The apparatus 300 may also be placed in the collectionorientation during stages of the post-collection processing of collectedtissue as described below. Accordingly, subsequent references herein tothe orientation of the apparatus 300, such as top, bottom, lower andupper, will refer to the collection orientation of FIG. 22. Asillustrated, the apparatus 300 has an apparatus height H_(A), anapparatus length L, and an apparatus depth (or width) D. The apparatus300 also include a suction port 302 and an inlet port 304. The suctionport 302 and inlet port 304 are disposed on the top of the apparatus 300when the apparatus 300 is in the collection orientation as illustratedin FIG. 22. In FIG. 22, and in certain other subsequent figures, theports are illustrated as having caps 328 thereon. Such caps 328 are usedto cover the various ports and may be removed and replaced as necessaryduring use of the apparatus 300.

The apparatus 300 includes a shell 306 and a lid 308. The shell 306 is aunitary bowl-like member where the only access into the interior, orcavity of the shell 306 is through the opening at the top of the shell306. As illustrated in FIG. 22, this opening at the top of the shell 306may be covered by the lid 308. The lid 308 and shell 306 may be rigid.The lid 308 and shell 306 are each preferably made of a clear polymericmaterial, such as a clarified polypropylene polymer composition, whichprovides low cellular adhesion and reasonable clarity. The lid 308 andshell 306 may be fabricated by injection molding. The lid 308 may beattached to the shell 306 in any appropriate manner, including snapping,clamping and/or gluing onto the shell 306. Together, the shell 306 andlid 308 form a container 322 with an internal containment volume 330(see FIG. 26 and accompanying discussion below) within the apparatus300. The internal containment volume 330 is the volume within the cavityof the shell 306 covered by the lid 308, and is the volume available fordisposing both hardware and material to be processed in the container322. This container 322 may have a container height H_(C). The shell 306may include a set of integral base supports 310 that may support theapparatus 300 in the collection orientation when the apparatus 300 isplaced on a horizontal surface. The apparatus height H_(A) is largerthan the container height H_(C) by the distance of projections above thetop of the container 322 for the inlet port 304, suction port 302, caps328 and other upward projecting features described below. The shell 306may be conveniently designed to efficiently fit within a centrifugebucket. The projections above the container height H_(C) may beconfigured so as not to interfere with operation of such a centrifuge.As seen in FIG. 22, the apparatus length L is equal to the containerlength and the apparatus depth is equal to the container depth (orwidth). As will be appreciated, the corresponding height, length anddepth dimensions of the internal containment volume 330 will equal theheight, length and depth dimensions of the container 322 less thecorresponding thicknesses of walls of the shell 306 and lid 308. Asshown in FIG. 22, some features may be integrally formed with the lid308. For example as shown in FIG. 22 the suction port 302 and the inletport 304 are integrally formed as a unitary fabricated piece with thelid 308. It should be appreciated that such features may be provided asseparate pieces and then assembled, such as by gluing or other means.For structural integrity, fabrication as a unitary piece is generallypreferred.

FIG. 23 shows another perspective view of the apparatus 300 with thecaps 328 to ports removed and with an installed handle 382. FIG. 24shows the apparatus 300 in the same orientation as in FIG. 23 with theshell 306 and handle 382 removed. With the shell 306 removed, a filter312 can be seen that is disposed within the internal containment volume330. The filter 312 may have a separation size in a range from 70microns to 400 microns. The filter is preferably made of a meshmaterial. The preferred mesh material is a nylon mesh. Also visiblewithin the internal containment volume 330 is a suction port conduit 314extending downward from the suction port 302. Additionally, asillustrated in FIG. 24, all components of the apparatus 300, except forthe shell 306, are interconnected to the lid 308. In this regard, thesubassembly shown in FIG. 24 may be assembled as shown and inserted intothe shell 306.

FIG. 25 shows another perspective view of the apparatus 300. FIG. 25shows the apparatus 300 in the same orientation as in FIG. 24 with boththe shell 306 and the filter 312 removed. With the filter 312 removed, aflow barrier skirt 324 extending downward from the lid 308 into theinternal containment volume 330 is visible. In an example, the flowbarrier skirt may extend between 5 mm and 50 mm downward from the lid308. The flow barrier skirt 324 may serve as an attachment point for thefilter 308 such that the filter 312 may be fixed relative to the lid308. The flow barrier skirt 324 may also serve to prevent material fromentering a tissue retention volume 332 (described below) and immediatelymoving through the filter 312 into the filtrate volume 334. The tissueretention volume 332 is that portion of the internal containment volume330 contained within the filter 312 and barrier skirt 324 below the lid308. The filtrate volume 334 is that portion of the internal containmentvolume 330 disposed outside of the filter 312 and barrier skirt 324.With the flow barrier skirt 324 in place, and material entering theinlet port 304 must at least move to below the lowest level of the flowbarrier skirt 324 before it is able to pass through the filter 312 intothe filtrate volume 334. The flow barrier skirt 324 may be part of afilter subassembly that includes the flow barrier skirt 324 and thefilter 312. This subassembly is mounted to the lid 308 with four screws326.

The filter 312 is asymmetric with respect to the lid 308 and shell 306in that it is configured to provide clearance between its left side (asviewed in FIG. 24) and the shell 306 for the suction port 302 andsuction port conduit 314. A portion of the filter 312 may be disposedabout (e.g., rest on or around) a portion of the suction port conduit314.

With the filter 312 removed (FIG. 25), a mixing device 316 can be seen.The mixing device 316 includes a rotatable shaft 318 and a set of mixingmembers 320. The axis of rotation of the rotatable shaft 318 may bethrough a central axis of the rotatable shaft 318. The mixing members320 are in the form of paddles extending outward from the rotatableshaft 318. Accordingly, when the rotatable shaft 318 is rotated, themixing members 320 will be rotated through the materials within thetissue retention volume 332 to aid in mixing the materials within theinternal containment volume 330, and in particular within the tissueretention volume 332. The rotatable shaft 318 extends from outside ofthe internal containment volume 330 through the lid 308 to the inside ofthe internal containment volume 330. As the rotatable shaft 318 isrotatable relative to the lid 308, the mixing members 320 fixed to therotatable shaft 318 are also rotatable relative to the lid 308. Therotatable shaft 318 may be made from a metal composition, such asstainless steel (e.g., grade 303, 304, or 316). Alternatively, therotatable shaft 318 may be made from a high-strength polymer compositionsuch as an Ultem™ resin product.

The rotatable shaft 318 may include a handle interface 380 (FIG. 24)that may provide an interface for the handle 382 (FIG. 23) to beinterconnected to the portion of the rotatable shaft 318 outside of theinternal containment volume 330. The handle interface 380 of FIG. 24 isin the form of a pair of parallel surfaces disposed about the portion ofthe rotatable shaft 318 outside of the internal containment volume 330′.The handle 382 has a mating pair of interior parallel surfacesconfigured such that when the handle 382 is placed over the handleinterface 380, turning the handle 382 will result in turning therotatable shaft 318 and the mixing device 316. Such an interface 380also allows for the handle 382 to be removed from and replaced on thehandle interface 380 as needed during use of the apparatus 300.

FIG. 26 is a side schematic view of the apparatus 300 showing the mixingdevice 316 and filter 312 within the shell 306. The internal containmentvolume 330 is the entire volume within the shell 306 and under the lid308. Together, the portions of the shell 306 and lid 308 that containthe internal containment volume 330 are a container 322 of the apparatus300. The filter 312 divides and separates the internal containmentvolume 330 of the container 322 into the tissue retention volume 332disposed inside the filter 312, and a filtrate volume 334 disposedwithin the shell 306 on the outside of the filter 312. The filtratevolume 334 is that portion of the internal containment volume 330 intowhich filtrate enters after passing through the filter 312 from thetissue retention volume 332.

Disposed within the internal containment volume 330 at the bottom of theshell 306, below a level 350 that is at or below the lowest extent ofthe filter 312 (and therefore also below the lowest extent of the tissueretention volume 332), is a collection volume 336, such that thecollection volume 336 occupies the lowermost portion of the filtratevolume 334 located below the lowest elevation of the tissue retentionvolume 332.

The shell 306 has a tapered wall portion 338 that defines a taperedportion 340 of the internal containment volume 330, such that thecross-sectional area of the tapered portion 340 of the internalcontainment volume 330 tapers with a reducing cross-sectional area in adirection toward bottom of the container 322. By tapering, it means thatthe cross-sectional area in a horizontal plane (assuming the apparatus300 is in the collection orientation) becomes smaller in the directionof the taper (e.g., a direction orthogonal to the horizontal plane). Thetapered portion 340 of the internal containment volume 330 occupies theportion of the internal containment volume 330 below a level 352 wherethe tapered wall portion 338 meets a straight wall portion 342 of theshell 306. The tapered wall portion 338 is shown as having a flat,uniform inclined wall surface. The incline angle of surfaces of thetapered wall portion need not be uniform from the top to the bottom ofthe tapered portion 340 of the internal containment volume 330, and mayvary from top to bottom with portions with different incline angles, andmay have a curved surface, provided that the cross-section area isreducing in the direction of the taper. Also, the tapered wall portion338 need not be uniform around the perimeter of the tapered portion 340of the internal containment volume 330. For example, in the embodimentin FIGS. 24-26, the tapered wall portion 338 has a steeper incline onthe ends than on the front or back of the apparatus 300.

The shell 306 may comprise an upper portion 344 having a first wallsurface portion 348 defining a corresponding upper portion 368 of theinternal containment volume 330. Substantially all of the first wallsurface portion 348 may have an incline relative to horizontal of atleast 75°. For example, substantially all of the first wall surfaceportion 348 may be substantially vertical (90° incline relative tohorizontal). The shell 306 may include a lower portion 346 located belowthe upper portion 344 and having a second wall surface portion 358defining a corresponding lower portion 360 of the internal containmentvolume 330. The lower portion 360 may include the tapered wall portion338 defining the tapered portion 340 of the internal containment volume330. Substantially all of the tapered wall portion 338 may preferablyhave an incline relative to horizontal in a range of from 30° to 60°,although other angles or curved surfaces may be used. The taperedportion 340 of the internal containment volume 330 may occupysubstantially the entire lower portion 360 of the internal containmentvolume 330. At least a first portion 362 of the filter 312 may bedisposed in the upper portion 368 of the internal containment volume 330and a second portion 364 of the filter 312 may be disposed in the lowerportion 360 of the internal containment volume 330. The tapered wallportion 338 may form a nadir 384 at its lowest elevation. The nadir 384may also be a nadir of the collection volume 336, the filtrate volume334, the container 322, and the internal containment volume 330.

The internal containment volume 330 may include an available processingvolume or “useable” volume 366 which may be the portion of the internalcontainment volume 330 that is usable and/or may normally be occupied bymaterials within the container 322 during normal use. For example, theavailable processing volume 366 may be the portion of the internalcontainment volume 330 below a level 356 that coincides with the bottomextension of a port through the lid 308 (such as a second suction port370 discussed below, see FIG. 22) and that is not occupied by portions(e.g., internal hardware) of the apparatus 300 within the internalcontainment volume 330, such as the mixing device 316, barrier skirt324, filter 312 and suction port conduit 314. The top of the availableprocessing volume may be at the elevation of the bottom extension of theinlet port 304, which may define a maximum fill level within theinternal containment volume 330.

The inlet port 304 in fluid communication with the tissue retentionvolume 332 through the lid 308 is configured for introducing tissuecomprising adipose directly into the tissue retention volume 332 duringa lipoplasty procedure. However, use of the apparatus 300 is not solimited, and the tissue may be introduced into the apparatus usingtissue previously collected in another container and transferred to theapparatus 300. An additional access port 372 in fluid communicationthrough the lid 308 with the tissue retention volume 332 provides anadditional route into the tissue retention volume 332, for example forintroduction of additives.

The suction port 302 is in fluid communication through the lid 308 withthe filtrate volume 334 via suction port conduit 314 extending from thesuction port 302 to within the tapered portion 340 of the internalcontainment volume 330 in the vicinity of the top of the collectionchamber 336. The suction port 302 is configured for connection to avacuum system, for example through connection of a suction conduitthrough which suction may be applied by a vacuum system to suction fromthe filtrate volume 334 material passing through the filter 312 from thetissue retention volume 332 into the filtrate volume 334.

Turning to FIGS. 27 and 28, the rotatable shaft 318 may include a filtercontact member 376 that is offset from an axis of rotation of therotatable shaft 318. A lower end of the filter contact member 376 maycontact a portion of the filter 312 as illustrated in FIG. 26. As therotatable shaft 318 is rotated, the filter contact member 376 may rotatein a circular path about the axis of rotation of the rotatable shaft 318remaining in contact with and moving along a portion of the filter 312.This contact may cause the filter 312 to deform and such deformationand/or the contact between the filter contact member 376 and filter 312may cause materials that may have adhered to the filter 312 in thisregion to become dislodged from the filter 312. Thus, the filter contactmember 376 may assist in keeping the filter from clogging and increasingthe effectiveness of the filter 312.

The rotatable shaft 318 may include a lumen 374 therethrough. The top ofthe lumen 374 is visible in FIG. 24 and the bottom of the lumen 374 isvisible in FIG. 28. The lumen 374 may have a distal end 386 (FIG. 28)within the tissue retention volume 332 and a proximal end 388 (FIG. 24)outside of the internal containment volume 330 and thus may allow accessto the tissue retention volume 332 therethrough. The lumen 374 may bedisposed along the central axis of the rotatable shaft 318. The lumen374 thus provides a conduit for accessing the internal containmentvolume 330. As further described below, the lumen 374 may provide accessfor removing processed material from the internal containment volume330. In that respect, the opening through the lid 308 through which therotatable shaft 308 extends acts as an extraction port through whichaccess is provided via the lumen 374 that passes through such opening.The apparatus 300 may include a plug 378, shown in FIG. 25 and not shownin FIG. 24 that may be placed in the proximal end 388 of lumen 374 toseal the lumen 374.

As illustrated in FIGS. 27 and 28, the apparatus 300 may include anoptional barrier member 390 (shown in FIG. 27, absent from FIG. 28). Thebarrier member 390 may be secured to the mixing device 316 via twoscrews 392 or by any other appropriate means such as snaps or by beingmolded integrally with the mixing members 320.

As shown in FIG. 29, in configurations where the barrier member 390 isnot present, a hypodermic needle 392 may be inserted through the lumen374 and may be advanced out of the distal end 386 of the lumen 374 andto pierce through the filter 312 to directly access the collectionvolume 336 (the volume under the line 410 in FIG. 29). Thus, without thebarrier member 390 present, the hypodermic needle 392 may be used toinject material into, or remove material from the collection volume 336.Additionally, as the axis of the lumen 374 is vertically oriented,access to the collection volume 336 using the hypodermic needle 392 isby downward vertical insertion into the lumen 374 from above thecontainer. Such vertical insertion coupled with the ability of theapparatus 300 to be placed on a flat surface in the collectionorientation, allows for user-friendly access to the collection volume336, and helps avoid complications that could compromise operations tocollect valuable processed material from the collection volume 336.

The hypodermic needle 392 may be interconnected to a syringe 394. Theproximal end 388 of the lumen 374 may include a tapered receptacleadapted to mate with a tapered tip of the syringe 394. In this regard,as shown in FIG. 29, the depth of penetration by the hypodermic needle392 into the collection volume 336 when the tapered tip of the syringe394 is in contact with the tapered receptacle of the lumen 374 may becontrolled by controlling the length of the hypodermic needle 392extending from the syringe 394. Additionally, the proximal end 388 ofthe lumen 374 may include a feature, such as a notch, to retain ano-ring (not shown) such that when the syringe 394 is positioned againstthe proximal end 388 of the lumen 374, the o-ring forms a seal betweenthe proximal end 388 of the lumen 374 and the syringe 394 (i.e., a sealthrough the o-ring between a wall surface in the tapered receptacle andan exterior wall surface of the tip of the syringe inserted into thetapered receptacle).

In configurations where the barrier member 390 is present, as shown inFIGS. 25 and 27, direct access from the lumen 374 to the collectionvolume 336 is prevented. Furthermore, the distance between the barriermember 390 and the distal end 386 of the lumen 374 may be selected toachieve a desired flow restriction through a gap between the distal end386 of the lumen 374 and the barrier member 390. For example, thedistance between the barrier member 390 and the distal end 386 of thelumen 374 may be between one and five millimeters. Such a distance maybe beneficial when the apparatus 300 is employed to perform a fat graftand the lumen 374 is used to remove tissue from the tissue retentionvolume 332. By maintaining an appropriate standoff between the barriermember 390 and the distal end 386 of the lumen 374, and by configuringthe barrier member with an appropriate areal extension beyond theperimeter of the opening of the distal end 376 of the lumen 374 (e.g.,the barrier member 390 is a large enough plate), potential for flowshort-circuiting to draw in air or other fluid through the filter 312from the filtrate volume 334 may be significantly reduced or avoidedwhen processed material (e.g., for a fat graft) is extracted by suctionthrough the lumen 374 into the syringe 379.

Turning to FIGS. 30A and 30B, the second suction port 370 includes atranslatable member 396 that may be translated up and down relative tothe lid 308 to vary the depth (elevation within the filtrate volume 334)at which material from the filtrate volume 334 is drawn through thesecond suction port 370. Examples of the various depths (elevations) atwhich the translatable member 396 may be positioned are illustrated inFIGS. 30A, 32, 33 and 35 and are discussed below in relation to methodsof using the apparatus 300. The fit between the translatable member 396and the opening through the lid 308 of the second suction port 370 issuch that the translatable member 396 may be readily translated up anddown to a desired level, while maintaining a tight enough fit to allow avacuum applied to the translatable member 396 to adequately drawmaterial out of the filtrate volume 334.

As illustrated in FIG. 31, the apparatus 300 may include a pre-filterunit 400 fluidly connected to the inlet port 304 via a length of tubing402. The pre-filter unit 400 may comprise within a housing a pre-filter,which may be in the form of a mesh screen with openings, for example,preferably in a range of from 0.5 millimeter to 2 millimeters. Thepre-filter unit 400 may be used to pre-filter tissue prior tointroduction into the tissue retention volume 332 of the apparatus 300.The tissue being pre-filtered may be supplied from a lipoplasty cannulaused during a lipoplasty procedure. Moreover, the suction port 302 ofthe apparatus 300 may be fluidly connected with a canister 206 via asuction conduit 208 similar to as illustrated in FIG. 6.

In general, the parts discussed with reference to the apparatus 300 maybe made from any appropriate biocompatible material. In particular, theshell 306 may be made from a biocompatible transparent polymer materialto allow inspection of the contents therein. Screws 326, 392 and therotatable shaft 318 may be made from metal, such as stainless steel.Other parts of the assembly 300 pictured in FIG. 22 may be made fromappropriate biocompatible polymers.

Various exemplary dimensions of one specific nonlimiting example of anapparatus 300 will now be described with reference to FIGS. 22 and 26.In this example, the apparatus 300 has apparatus height H_(A) of about157 mm, an apparatus length L of about 145 millimeters, and an apparatusdepth D of about 126 millimeters. The containment volume height H_(C) isabout 124 millimeters. The example has an available processing volume366 of about 760 milliliters and a collection volume of about 23milliliters. The portion of the tissue retention volume 332 thatcoincides with the available processing volume 366 is about 580milliliters.

In a method for processing tissue from lipoplasty procedure using theapparatus 300, the tissue is processed within the internal containmentvolume 330 to prepare within the apparatus 300 a concentrated productcomprising at least one target component, or at least one targetmaterial, from the tissue. For such a procedure, the barrier member 390is not present within the apparatus 300. Many features of the previouslydiscussed methods may also be employed in the current method whereappropriate. Such features include, inter alia, multiple washings,shaking, heating, and centrifuging as previously described. Returning tothe present method, the tissue is introduced into the tissue retentionvolume 332 through the inlet port 304. The tissue may be pre-filteredusing pre-filter unit 400 prior to being introduced into the tissueretention volume 332. The method may comprise washing tissue in theinternal containment volume 330 with a wash liquid. Optionally, thewashing may include centrifuging the apparatus 300. After washing, themethod may comprise digesting tissue within the internal containmentvolume 330. After the digestion, the method may include centrifuging theapparatus 300 to prepare in the collection volume 336 a concentrateproduct comprising at least one target component. For example theconcentrate product may comprise, or may consist essentially of, stromalvascular fraction from adipose tissue, and a target component may bestem cells from adipose tissue.

During the washing, the wash liquid may be added to the internalcontainment volume 330 to contact tissue within the tissue retentionvolume 332 and with at least a portion, preferably a majority, and morepreferably most, of the wash liquid passing through the filter 312 intothe filtrate volume 334. The addition of the tissue to the internalcontainment volume 330 may occur simultaneously with the wash liquidbeing removed from the filtrate volume 334 via vacuum applied to thesuction port 302. In this regard, a volume of tissue larger than theinternal containment volume 330 may be introduced into the internalcontainment volume 330 during the performance of the method. Moreover,the removal of wash liquid may continue after the introduction of tissueinto the internal containment volume 330 has stopped.

The wash liquid may wash one or more components from the tissue whileretaining washed tissue in the tissue retention volume 332. The washedtissue may be retained in the tissue retention volume 332 by the filter312. Wash liquid passing into the filtrate volume 334 may be removedfrom the filtrate volume 334, along with any component or componentswashed from the tissue. Optionally, after adding the wash liquid, theapparatus 300 may be centrifuged to facilitate a high degree ofseparation of the wash liquid from the tissue retained in the tissueretention volume 332. Next, the wash liquid may be removed from thefiltrate volume 334 by suctioning through the suction port 302 of theapparatus 300. The washing may include multiple wash stages. During thewashing, the mixing device 316 may be rotated by rotating the handle 382to assist in the washing process.

During the digestion, an enzyme, such as for example collagenase, may beadded to the internal containment volume 330 through the additionalaccess port 372 or through the inlet port 304. During the digesting, themixing device 316 may be rotated to assist in the digesting process.

After adding the enzyme, the digesting may comprise agitating contentsof the containment volume of the apparatus 300 for a time and at atemperature sufficient for the digestion to proceed to an extent tosignificantly release the target component, or material, in the desiredform capable of passing through the filter 312. The agitating mayinvolve any method to agitate contents of the internal containmentvolume 330, including for example one or both of: (a) shaking theapparatus 300 to agitate the contents within the apparatus 300 and (b)mixing the contents within the apparatus 300 by rotating the mixingdevice 316 using the handle 382.

Post-digestion centrifuging promotes separation of the target componentfrom the digested tissue and passage of the target component through thefilter 312 for collection in the collection volume 336. The targetcomponent may include stem cells from adipose tissue. As illustrated inFIG. 32, multiple material phases may collect within the filtrate volume334. The first (bottom) material phase may be a layer of red blood cells428 located in the region of the filtrate volume 334 below the line 408.This volume below the line 408 occupies a bottom portion of thecollection volume 336. The second material phase may be a stromalvascular fraction layer 430 from adipose tissue and may be located inthe region of the filtrate volume 334 below the line 412 and above theline 408. As will be appreciated, the red blood cell layer 428 and thestromal vascular fraction layer may not be divided by a sharp line, andthe blood cell layer 428 may grade into the lower portion of the stromalvascular fraction layer 430. This volume below the line 412 and abovethe line 408 also occupies a portion of the collection volume 336. Athird material phase may be an aqueous layer 432 that occupies theregion of the filtrate volume 334 below the line 406 and above the line412. A fourth material phase may be a disaggregated adipose layer 434that occupies the region of the filtrate volume 334 below the line 414and above the line 406. A fifth material phase may be an oil layer 436that occupies the region of the filtrate volume 334 below the line 416and above the line 414. The separated phase layers as shown are providedto illustrate relative positioning and are not intended to represent anactual scale of the relative sizes of the phases, except that the redblood cell layer 428 and stromal vascular fraction layer 430 arecontained within the collection volume 336 and the other layers extendabove the collection volume.

The translatable member 396 of the second suction port 370 may beemployed to first remove the oil layer 436, then to remove thedisaggregated adipose layer 434, and then to remove the aqueous layer432. As illustrated in FIG. 32, the translatable member 396 may bepositioned such that the end of the translatable member 396 is disposedwithin the oil layer 436. Suction applied to the translatable member 396will remove the As fluid is removed, the translatable member may belowered to remove additional fluid down to a desired level, which may beremoval of all or most of layers 436, 434 and 432. For example, once theoil layer 436 has been removed, the translatable member 396 may belowered into the disaggregated adipose layer 434 and then the aqueouslayer 432 for sequential removal of these layers. FIG. 33 illustratesthe aqueous layer 432 partially removed (after already removing the toplayers 436 and 434 such that the top of the aqueous layer 432 is at line418). As another example, the translatable member 396 may be initiallyinserted to the position shown in FIG. 33 and suction applied until aportion of the aqueous layer 432 is removed and also the disaggregatedadipose layer 434 and oil layer 436 are removed above line 418,resulting in the arrangement of FIG. 33.

Once fully inserted into the filtrate volume 334, the translatablemember 396 may not be operable to remove a portion of the aqueous layer432 while the apparatus 300 is in the collection orientation.Accordingly, a user may gently tilt the apparatus 300 as illustrated inFIG. 34 to further remove the aqueous layer 432. As illustrated, thestromal vascular fraction 430 below the line 412 may form a pellet whichmay retain its position as the apparatus 300 is tilted. This attributeof the pellet allows the apparatus 300 to be tilted such that theaqueous layer 432 flows toward the translatable member 396 disposedproximate to the interface between the tapered wall portion 338 of theshell 306 and the straight wall portion 342 of the shell 306 asillustrated by line 420 in FIG. 34. Such tilting can allow suction to beapplied to the aqueous layer 432 without the suction substantiallyaffecting the stromal vascular fraction 430. Once the aqueous layer 432has been satisfactorily removed, the apparatus 300 may be returned toits collection orientation, as shown in FIG. 35, for removal of thestromal vascular fraction 430 from the collection volume 336.

Next, the hypodermic needle 394 may be inserted into the collectionvolume 336 as illustrated in FIG. 29 and a diluent fluid may be injectedinto the collection volume 336 such that the diluent fluid, stromalvascular fraction 430 and the layer of red blood cells 428 togetheroccupy at least a portion of the collection volume 336 under line 410,and are preferably limited to the collection volume 336 and do notoccupy space above line 410. After injection of the diluent fluid, auser may gently tap the apparatus 300 against a hard surface to causethe diluent fluid to mix with the stromal vascular fraction and thelayer of red blood cells. A second hypodermic needle may then beinserted through the lumen 374 and the diluent/stromal vascularfraction/red blood cell mixture may be removed from the apparatus 300.

In a method for processing tissue from a lipoplasty procedure using theapparatus 300, the tissue is collected into and processed within theinternal containment volume 330 to prepare within the apparatus 300 afat graft composition for reintroduction into a patient. For such aprocess, the barrier member 390 will be present within the apparatus 300and positioned as shown in FIG. 25. The method may comprise collectingand washing adipose tissue in the internal containment volume 330 in amanner similar to that discussed above with respect to the preparationof the stromal vascular fraction. The washing may or may not includeoptional centrifuging. In a variation, the apparatus 300 may bespecifically configured such that it cannot be centrifuged using thetype of centrifuge that the apparatus 300 is typically inserted into(e.g., the centrifuge 244 of FIG. 20). Such incompatibility may beachieved, for example, by adding ribs or extensions to the lid 308and/or shell 306 such that the apparatus 300 cannot fit into thecentrifuge 244. After washing the present method may include adding anadditive to the washed tissue in the tissue retention volume 332. Theadditive may be added through the additional access port 372 or throughthe inlet port 304. The additive may comprise an active ingredient fordelivery to the patient in a fat graft. The mixing device 316 may berotated to assist in distributing the additive throughout the tissuewithin the tissue retention volume 332.

Once the fat graft composition is prepared, it may be removed from thetissue retention volume 332 by applying suction to the proximal end ofthe lumen 374. Such applied suction will pull the fat graft compositionmaterial through the lumen 374 and out of the tissue retention volume332. During such suction, the barrier member 390, by being interposedbetween the distal end of the lumen 374 and the filter 312 will preventthe filter 312 from contacting the lumen 374 which could interfere withthe contents of the tissue retention volume 332 being drawn into thelumen 374. Moreover, by having the barrier member 390 a fixedpredetermined distance from the distal end 386 of the lumen 374, thematerial within the tissue retention volume 332 must flow through therestricted space between the barrier member 390 and the distal end 386of the lumen 374. The restricted space and areal extent of the barriermember may serve to limit the flow rate of material moving from thetissue retention volume 332 into the lumen 374.

The foregoing discussion of the invention and different aspects thereofhas been presented for purposes of illustration and description. Theforegoing is not intended to limit the invention to only the form orforms specifically disclosed herein. Consequently, variations andmodifications commensurate with the above teachings, and the skill orknowledge of the relevant art, are within the scope of the presentinvention. The embodiments described hereinabove are further intended toexplain best modes known for practicing the invention and to enableothers skilled in the art to utilize the invention in such, or other,embodiments and with various modifications required by the particularapplications or uses of the present invention. It is intended that theappended claims be construed to include alternative embodiments to theextent permitted by the prior art. Although the description of theinvention has included description of one or more possibleimplementations and certain variations and modifications, othervariations and modifications are within the scope of the invention,e.g., as may be within the skill and knowledge of those in the art afterunderstanding the present disclosure. It is intended to obtain rightswhich include alternative embodiments to the extent permitted, includingalternate, interchangeable and/or equivalent structures, functions,ranges or steps to those claimed, whether or not such alternate,interchangeable and/or equivalent structures, functions, ranges or stepsare disclosed herein, and without intending to publicly dedicate anypatentable subject matter. Furthermore, any feature described or claimedwith respect to any disclosed implementation may be combined in anycombination with one or more of any other features of any otherimplementation or implementations, to the extent that the features arenot necessarily technically compatible, and all such combinations arewithin the scope of the present invention.

The terms “comprising”, “containing”, “including” and “having”, andgrammatical variations of those terms, are intended to be inclusive andnonlimiting in that the use of such terms indicates the presence of somecondition or feature, but not to the exclusion of the presence also ofany other condition or feature. The use of the terms “comprising”,“containing”, “including” and “having”, and grammatical variations ofthose terms in referring to the presence of one or more components,subcomponents or materials, also include and is intended to disclose themore specific embodiments in which the term “comprising”, “containing”,“including” or “having” (or the variation of such term) as the case maybe, is replaced by any of the narrower terms “consisting essentially of”or “consisting of” or “consisting of only” (or the appropriategrammatical variation of such narrower terms). For example, the astatement that some thing “comprises” a stated element or elements isalso intended to include and disclose the more specific narrowerembodiments of the thing “consisting essentially of” the stated elementor elements, and the thing “consisting of” the stated element orelements. Examples of various features have been provided for purposesof illustration, and the terms “example”, “for example” and the likeindicate illustrative examples that are not limiting and are not to beconstrued or interpreted as limiting a feature or features to anyparticular example. The term “at least” followed by a number (e.g., “atleast one”) means that number or more than that number. The term at “atleast a portion” means all or a portion that is less than all. The term“at least a part” means all or a part that is less than all.

What is claimed is:
 1. An apparatus useful for collection of humanbiological material and post-collection processing of collectedmaterial, the apparatus being orientable in a collection orientation, asoriented in the collection orientation the apparatus comprising: afilter; a container having an internal containment volume, the internalcontainment volume comprising: a tissue retention volume and a filtratevolume separated by the filter; a collection volume within the filtratevolume, the collection volume having a bottom elevation corresponding toa bottom elevation of the filtrate volume and a top elevation that islower than the bottom elevation of the tissue retention; a taperedportion that tapers in a downward direction with at least a portion ofthe tapered portion being located above the collection volume; an inletport in fluid communication with the tissue retention volume andconfigured for introducing human biological material comprising adiposedirectly into the tissue retention volume; a suction port in fluidcommunication with the filtrate volume and providing access to thefiltrate volume for suctioning from the filtrate volume componentspassing through the filter from the tissue retention volume to thefiltrate volume; and a mixing device disposed at least in part withinthe tissue retention volume for mixing contents within the tissueretention volume.
 2. An apparatus according to claim 1, wherein themixing device comprises a rotatable shaft extending from outside of theinternal containment volume to inside of the internal containmentvolume. 3-5. (canceled)
 6. An apparatus according to claim 2, comprisinga lumen extending through the shaft and having a proximal end locatedoutside of the internal containment volume and a distal end locatedwithin the internal containment volume, thereby providing access fromoutside of the internal containment volume to inside of the internalcontainment volume.
 7. (canceled)
 8. An apparatus according to eitherclaim 6, wherein the shaft is rotatable about an axis that extendsthrough the collection volume.
 9. An apparatus according to claim 8,wherein the lumen is aligned with the axis. 10-15. (canceled)
 16. Anapparatus according to claim 6, wherein the apparatus is configured foradvancing a hypodermic needle through the lumen and out of the distalend of the lumen to access the collection volume with an advancing tipof the hypodermic needle.
 17. An apparatus according to claim 16,wherein the distal end of the lumen is located in the tissue retentionvolume above a portion of the filter, so that the advancing tip of ahypodermic needle pierces the filter when the tip of a hypodermic needleexits the distal end of the lumen and is advanced from the distal end ofthe lumen into the collection volume.
 18. An apparatus according toclaim 16, wherein the collection volume includes a nadir, and an axis ofthe lumen is aligned so that the tip of a hypodermic needle exiting thedistal end of the lumen may be advanced to the vicinity of the nadir ofthe collection volume.
 19. An apparatus according to claim 2, whereinthe mixing device comprises at least one mixing member disposed in thetissue retention volume and connected with the shaft, wherein the atleast one mixing member moves through the tissue retention volume whenthe shaft is rotated.
 20. (canceled)
 21. An apparatus according to claim2, wherein when the mixing device comprises a filter contact member thatmoves when the shaft is rotated, the filter contact member contactingthe filter at least periodically when the shaft is rotated. 22.(canceled)
 23. An apparatus according to claim 1, wherein the suctionport is in fluid communication with the tapered portion of the internalcontainment volume through a conduit providing fluid communication fromthe suction port to a location within the filtrate volume and within thetapered portion of the internal containment volume. 24-25. (canceled)26. An apparatus according to claim 23, wherein: the suction port is afirst suction port, the conduit is a first conduit and the locationwithin the filtrate volume is a first location within the filtratevolume; the apparatus comprises a second suction port through whichcomponents passing through the filter from the tissue retention volumeto the filtrate volume may be suctioned from the filtrate volume; andthe apparatus comprises a second conduit extending from the secondsuction port to a second location within the filtrate volume, the secondconduit being translatable through the second suction port to adjust theelevation of the second location within the filtrate volume. 27-34.(canceled)
 35. An apparatus according to claim 1, wherein all access tothe internal containment volume is through access ports wherein eachsaid access port is configured for access through the said access portfrom above the container. 36-37. (canceled)
 38. An apparatus accordingto claim 1, wherein the container comprises: a fluid containment shellwith an internal cavity portion forming at least a part of the internalcontainment volume, the internal cavity portion being open to above; anda lid attached to the shell and disposed to cover from above theinternal cavity portion; wherein, the suction port and the inlet porteach passes through the lid. 39-40. (canceled)
 41. An apparatusaccording to claim 38, wherein the filter is suspended from the lid andthe mixing device is supported by the lid and extends verticallydownward from the lid into the tissue retention volume. 42-53.(canceled)
 54. An apparatus according to claim 1, wherein the internalcontainment volume has an available processing volume in a range of from100 cubic centimeters to 1000 cubic centimeters. 55-60. (canceled) 61.An apparatus according to claim 54, wherein the apparatus is containablewithin a first envelope volume defined by a rectangular cuboid having alength dimension of no more than 16 centimeters, a depth dimension of nomore than 15 centimeters and a height dimension of no more than 18centimeters. 62-63. (canceled)
 64. An apparatus according to claim 1,wherein the filter has a separation size in a range of from 70 micronsto 400 microns. 65-69. (canceled)
 70. An apparatus according to claim 1,comprising human biological material disposed within the internalcontainment volume.
 71. An apparatus according to claim 1, comprising,disposed within the collection volume, a stromal vascular fraction fromadipose tissue.
 72. An apparatus according to claim 1, wherein theapparatus is sterilized and packaged within a hermetic enclosure. 73-78.(canceled)
 79. An apparatus useful for collection of human biologicalmaterial, and post-collection processing of collected material, theapparatus being orientable in a collection orientation, as oriented inthe collection orientation the apparatus comprising: a filter; acontainer having an internal containment volume having a volume in arange of from 100 cubic centimeters to 1300 cubic centimeters, theinternal containment volume comprising: a tissue retention volume and afiltrate volume separated by the filter; a collection volume within thefiltrate volume, the collection volume having a bottom elevationcorresponding to a bottom elevation of the filtrate volume and a topelevation that is lower than the bottom elevation of the tissueretention volume; a tapered portion that tapers in a downward directionwith at least a portion of the tapered portion being located above thecollection volume; an inlet port in fluid communication with the tissueretention volume and configured for introducing human biologicalmaterial comprising adipose directly into the tissue retention volumeduring a lipoplasty procedure; a suction port in fluid communicationwith the filtrate volume and providing access to the filtrate volume forsuctioning from the filtrate volume components passing through thefilter from the tissue retention volume to the filtrate volume; and anextraction port in fluid communication with the internal containmentvolume and configured for removing processed biological material fromthe internal containment volume; wherein each of the inlet port, thesuction port and the extraction port is configured for accesstherethrough from above the container into the internal containmentvolume. 80-127. (canceled)
 128. An assembly comprising: the apparatusaccording to claim 1; and a centrifuge; wherein the apparatus isreceived in the centrifuge for centrifugation of the apparatus. 129-146.(canceled)
 147. A method for processing human biological materialcomprising adipose within the internal containment volume of anapparatus according to claim 1, the method comprising: washing the humanbiological material with a wash liquid, the washing comprising addingthe wash liquid to the internal containment volume to contact the humanbiological material within the tissue retention volume and passingthrough the filter and removing from the filtrate volume at least aportion of the wash liquid along with one or more components washed fromthe human biological material while retaining washed human biologicalmaterial in the tissue retention volume. 148-151. (canceled)
 152. Amethod according to claim 147, comprising adding an additive to thewashed human biological material in the tissue retention volume. 153.(canceled)
 154. A method according to claim 152, wherein the additivecomprises an ingredient for delivery to a patient in a fat graft.
 155. Amethod according to claim 154, wherein the additive comprises a memberselected from the group consisting of hormones and stem cells.
 156. Amethod according to claim 147, comprising removing from the tissueretention volume at least a portion of the washed human biologicalmaterial for use in a fat graft.
 157. A method according to claim 147,comprising digesting material within the containment volume, thedigesting material comprising adding enzyme to the containment volume tocontact at least a portion of the washed human biological materialwithin the tissue retention volume, the enzyme being of a type capableof breaking down a portion of the washed human biological material torelease target material in a form capable of passing through the filter.158. (canceled)
 159. A method according to claim 157, comprising afterthe digesting, centrifuging the apparatus to prepare in the collectionvolume a concentrate product comprising the target material, the targetmaterial comprising stromal vascular cells. 160-162. (canceled)
 163. Amethod according to claim 159, comprising: after the centrifuging,selectively removing the concentrate product from the filtrate volume;and wherein after the centrifuging and prior to the selectivelyremoving, the concentrate product is contained in a bottom separatedlayer or layers disposed within the collection volume, and theselectively removing comprises removing from the filtrate volume otherseparated layers within the filtrate volume disposed above the bottomseparated layer or layers after the centrifuging.
 164. A methodaccording to claim 163, wherein the apparatus comprises a second suctionport, and the removing from the filtrate volume other separated layerswithin the filtrate volume disposed above the bottom separated layer orlayers comprises: removing material of the other separated layersthrough the second suction port.
 165. A method according to claim 163,wherein the removing from the filtrate volume other separated layerswithin the filtrate volume disposed above the bottom separated layer orlayers comprises: tilting the container and suctioning material of theother layers from a laterally distant location that is laterally distantfrom an outer edge of the concentrate product, wherein during thesuctioning the concentrate product remains in a pellet located adjacentthe bottom of the collection volume. 166-167. (canceled)
 168. A methodaccording to claim 163, comprising after the removing from the filtratevolume separated layers within the filtrate volume disposed above thebottom separated layer or layers: diluting the bottom layer with adiluent fluid to prepare a diluted concentrate product, and removing thediluted concentrate product from the filtrate volume. 169-188.(canceled)